45 results on '"Hattori, Shunji"'
Search Results
2. Silibinin relieves UVB-induced apoptosis of human skin cells by inhibiting the YAP-p73 pathway
- Author
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Liu, Wei-wei, Wang, Fang, Li, Can, Song, Xiao-yu, Otkur, Wuxiyar, Zhu, Yu-ying, Hayashi, Toshihiko, Mizuno, Kazunori, Hattori, Shunji, Fujisaki, Hitomi, and Ikejima, Takashi
- Abstract
Excessive exposure to UVB induces skin diseases. Silibinin, a flavonolignan used for treating liver diseases, is found to be effective against UVB-caused skin epidermal and dermal cell damage. In this study we investigated the molecular mechanisms underlying. Human nonmalignant immortalized keratinocyte HaCaT cells and neonatal human foreskin fibroblasts HFFs were exposed to UVB irradiation. We showed that pre-treatment with silibinin dose-dependently decreased UVB-induced apoptosis of HaCaT cells. Furthermore, we showed that silibinin treatment inhibited nuclear translocation of YAP after UVB irradiation. Molecular docking analysis and DARTS assay confirmed the direct interaction of silibinin with YAP. Silencing YAP by siRNA had no influence on the survival of HaCaT cells, whereas inhibiting classical YAP-TEAD signaling pathway by siRNA targeting TEAD1 or its pharmaceutical inhibitor verteporfin further augmented UVB-induced apoptosis, suggesting that YAP-TEAD pathway was prosurvival, which did not participate in the protective effect of silibinin. We then explored the pro-apoptotic YAP-p73 pathway. p73 was upregulated in UVB-irradiated cells, but reduced by silibinin cotreatment. The mRNA and protein levels of p73 target genes (PML, p21and Bax) were all increased by UVB but decreased by silibinin co-treatment. Inhibiting p73 by using siRNA reduced UVB-induced apoptosis, suggesting that downregulation of p73 was responsible for the cytoprotective effect of silibinin. In HFFs, the upregulated YAP-p73 pathway by UVB irradiation was also suppressed by silibinin. Collectively, YAP-p73 pathway is a major cause of the death of UVB-exposed epidermal HaCaT cells and dermal HFFs. Silibinin directly inhibits YAP-p73 pathway, exerting the protective action on UVB-irradiated skin cells.
- Published
- 2021
- Full Text
- View/download PDF
3. Role of Glycosyltransferase 25 Domain 1 in Type I Collagen Glycosylation and Molecular Phenotypes
- Author
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Terajima, Masahiko, Taga, Yuki, Sricholpech, Marnisa, Kayashima, Yukako, Sumida, Noriko, Maeda, Nobuyo, Hattori, Shunji, and Yamauchi, Mitsuo
- Abstract
Glycosylation in type I collagen occurs as O-linked galactosyl- (G-) lesser and glucosylgalactosyl-hydroxylysine (GG-Hyl); however, its biological significance is still not well understood. To investigate the function of this modification in bone, we have generated preosteoblast MC3T3-E1 (MC)-derived clones, short hairpin (Sh) clones, in which Glt25d1gene expression was stably suppressed. In Sh clones, the GLT25D1 protein levels were markedly diminished in comparison to controls (MC and those transfected with the empty vector). In Sh collagen, levels of both G- and GG-Hyl were significantly diminished with a concomitant increase in the level of free-Hyl. In addition, the level of immature divalent cross-links significantly diminished while the level of the mature trivalent cross-link increased. As determined by mass spectrometric analysis, seven glycosylation sites were identified in type I collagen and the most predominant site was at the helical cross-linking site, α1–87. At all of the glycosylation sites, the relative levels of G- and GG-Hyl were markedly diminished, i.e., by ∼50–75%, in Sh collagen, and at five of these sites, the level of Lys hydroxylation was significantly increased. The collagen fibrils in Sh clones were larger, and mineralization was impaired. These results indicate that GLT25D1 catalyzes galactosylation of Hyl throughout the type I collagen molecule and that this modification may regulate maturation of collagen cross-linking, fibrillogenesis, and mineralization.
- Published
- 2019
- Full Text
- View/download PDF
4. Semi-Nondestructive Certification of Crocodilian Leather by LC–MS Detection of Collagen Marker Peptides.
- Author
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Kumazawa, Yuki, Hattori, Shunji, and Taga, Yuki
- Published
- 2019
- Full Text
- View/download PDF
5. The C‐terminal segment of collagenase in Grimontia hollisae binds collagen to enhance collagenolysis.
- Author
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Tanaka, Keisuke, Teramura, Naoko, Hayashida, Osamu, Iijima, Katsumasa, Okitsu, Teru, and Hattori, Shunji
- Subjects
C-terminal residues ,COLLAGENASES ,COLLAGEN ,RECOMBINANT proteins ,FLUORESCEIN isothiocyanate - Abstract
The collagenase secreted by Grimontia hollisae strain 1706B is a 74 kDa protein that consists of two parts: the catalytic module and a C‐terminal segment that includes the bacterial pre‐peptidase C‐terminal domain. Here, we produced a recombinant C‐terminal segment protein and examined its ability to bind collagen and other characteristics as compared with collagen‐binding domains (CBDs) derived from Hathewaya histolytica (Clostridium histolyticum) collagenases; these CBDs are the only ones thus far identified in bacterial collagenases. We found that the C‐terminal segment binds to collagen only when the collagen is in its triple‐helical conformation. Moreover, the C‐terminal segment and the CBDs from H. histolytica have comparable characteristics, including binding affinity to type I collagen, substrate spectrum, and binding conditions with respect to salt concentration and pH. However, the C‐terminal segment has a completely different primary structure from those of the CBDs from H. histolytica. As regards secondary structure, in silico prediction indicates that the C‐terminal segment may be homologous to those in CBDs from H. histolytica. Furthermore, we performed collagenase assays using fluorescein isothiocyanate‐labeled type I collagen to show that the C‐terminal segment positively contributes to the collagenolytic activity of the 74 kDa collagenase from G. hollisae. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
6. Collagen-derived X-Hyp-Gly-type tripeptides promote differentiation of MC3T3-E1 pre-osteoblasts.
- Author
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Taga, Yuki, Kusubata, Masashi, Ogawa-Goto, Kiyoko, Hattori, Shunji, and Funato, Noriko
- Abstract
Oral ingestion of collagen hydrolysate leads to significantly high blood levels of various hydroxyproline (Hyp)-containing oligopeptides, such as Pro-Hyp that has been reported to promote osteoblast differentiation. Using ginger protease, we recently developed a novel type of collagen hydrolysate enriched with X-Hyp-Gly-type tripeptides, which were efficiently absorbed into the blood after oral administration of the ginger-degraded collagen hydrolysate. In the present study, we demonstrated that X-Hyp-Gly promoted the differentiation of MC3T3-E1 pre-osteoblasts more effectively than Pro-Hyp, as evidenced by increases in alkaline phosphatase activity and mineralization. In addition, collagen secretion was enhanced by Ala-Hyp-Gly and Leu-Hyp-Gly, particularly at an early stage of differentiation. We detected the uptake of Ala-Hyp-Gly and Leu-Hyp-Gly as well as Pro-Hyp into MC3T3-E1 cells. These results suggest that the collagen hydrolysate prepared using ginger protease can be used as a functional food for the prevention and treatment of bone diseases, such as osteoporosis. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
7. Semi-Nondestructive Certification of Crocodilian Leather by LC–MS Detection of Collagen Marker Peptides
- Author
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Kumazawa, Yuki, Hattori, Shunji, and Taga, Yuki
- Abstract
Leather produced from crocodile, alligator, and caiman skin is widely used in the fashion industry. Crocodilian leather is generally more expensive than mammalian leather, and the value greatly differs even between the crocodilian species. However, inappropriate labeling of the animal source on leather products sometimes arises from accidental or fraudulent substitution, which is difficult to unambiguously detect by existing methods. In the present study, animal source identification of crocodilian leather was carried out using type I collagen-derived marker peptides generated after dechroming, heat denaturation, and trypsin digestion. Definitive discrimination between the three crocodilian species and also a related species, lizard, was achieved based on the detection patterns of selected six marker peptides, determined by LC–MS. Furthermore, powdering of the leather samples enabled a reduction in the sample amount required and allowed the elimination of the dechroming step. Approximately 100 μg of powder was taken from commercial leather watch straps by filing, resulting in only slight damage to the undersides of the straps. The animal sources of the crocodilian products and also a crocodile-embossed calf product were successfully identified using a combination of the crocodilian marker peptides and previously established mammalian marker peptides. This semi-nondestructive species identification method is not only useful for certification of leather products but also for monitoring of international trade of leather and skin.
- Published
- 2019
- Full Text
- View/download PDF
8. A novel LC–MS method using collagen marker peptides for species identification of glue applicable to samples with multiple animal origins
- Author
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Kumazawa, Yuki, Taga, Yuki, Takashima, Miho, and Hattori, Shunji
- Abstract
Animal glues derived from collagen-rich tissue are widely used as adhesives and binders in artworks. Identification of the animal source of glue is important not only to understand the artistic technique and historical background, but also to conduct appropriate treatment for restoration and conservation of artworks. Recently, mass spectrometric approaches have been used for species identification of glue. However, it appears that the existing methods were not applied to samples containing multiple animal sources, and the range of target species has been limited because of incomplete sequence information of collagen in public databases. In the present study, we established a novel method for discrimination of glues from eight animals applicable to samples with multiple animal origins. Trypsin-digested glue samples were analyzed by LC–MS in multiple reaction monitoring mode, and the animal source was determined based on the detection patterns of 12 type I collagen-derived marker peptides. Using the database-independent method, we successfully identified the animal source of commercial products and found the declared species for some glues to be incorrect. In addition, some products were identified to contain two different animal origins. Relative abundance of the animal origins in several impure glues was estimated using the marker peptides, which helped to speculate the reason for the detection of multiple species. We analyzed a painting (The Harvestby Camille Pissarro, 1882) and clarified that glues used in the ground and size layers of the canvas were derived from cattle (~ 65%) and sheep (~ 35%).
- Published
- 2018
- Full Text
- View/download PDF
9. Production of a novel wheat gluten hydrolysate containing dipeptidyl peptidase-IV inhibitory tripeptides using ginger protease.
- Author
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Taga, Yuki, Hayashida, Osamu, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Subjects
GLIADINS ,GLUTELINS ,CD26 antigen - Abstract
Wheat gluten is a Pro-rich protein complex comprising glutenins and gliadins. Previous studies have reported that oral intake of enzymatic hydrolysates of gluten has beneficial effects, such as suppression of muscle injury and improvement of hepatitis. Here, we utilized ginger protease that preferentially cleaves peptide bonds with Pro at the P2position to produce a novel type of wheat gluten hydrolysate. Ginger protease efficiently hydrolyzed gluten, particularly under weak acidic conditions, to peptides with an average molecular weight of <600 Da. In addition, the gluten hydrolysate contained substantial amounts of tripeptides, including Gln-Pro-Gln, Gln-Pro-Gly, Gln-Pro-Phe, Leu-Pro-Gln, and Ser-Pro-Gln (e.g. 40.7 mg/g at pH 5.2). These gluten-derived tripeptides showed high inhibitory activity on dipeptidyl peptidase-IV with IC50values of 79.8, 70.9, 71.7, 56.7, and 78.9 μM, respectively, suggesting that the novel gluten hydrolysate prepared using ginger protease can be used as a functional food for patients with type 2 diabetes. Production of DPP-IV inhibitory tripeptides from gluten using ginger protease. [ABSTRACT FROM PUBLISHER]
- Published
- 2017
- Full Text
- View/download PDF
10. Development of immersion vaccine for bacterial cold-water disease in ayu Plecoglossus altivelis.
- Author
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Nakayama, Hitoshi, Mori, Mariko, Takita, Teisuke, Yasukawa, Kiyoshi, Tanaka, Keisuke, Hattori, Shunji, Aikawa, Hideaki, Hasegawa, Osamu, Okamura, Takashi, Takegami, Kentarou, Motokawa, Shogo, Kuwahara, Masakazu, and Amano, Kenichi
- Subjects
AYU ,FISH diseases ,BACTERIAL vaccines ,VACCINATION - Abstract
Flavobacterium psychrophilum(F. psychrophilum) is the causative agent of bacterial cold-water disease (BCWD) that occurs in ayuPlecoglossus altivelis. Formalin-killed cell ofF. psychrophilumhas long been studied as an immersion vaccine for BCWD. In this study, we explored the possibility ofF. psychrophilumcollagenase (fpcol) for use as the immersion vaccine. BCWD convalescent ayu sera contained specific IgM antibodies against somaticF. psychrophilumand fpcol, meaning that fpcol is a promising antigen for the vaccine development. The recombinant fpcol was successfully expressed inEscherichia coliandBrevibacillus chosinensis(B. chosinensis). The culture supernatant of theB. chosinensiswas used as an immersion vaccine solution. The vaccinated ayu were then challenged by soaking intoF. psychrophilumculture. In two experimental groups, the relative percentages of survivals were 63 and 38%, respectively, suggesting that fpcol is promising as the immersion vaccine for ayu-BCWD. [ABSTRACT FROM PUBLISHER]
- Published
- 2017
- Full Text
- View/download PDF
11. The cleavage site preference of the porcine pepsin on the N-terminal α1 chain of bovine type I collagen: a focal analysis with mass spectrometry.
- Author
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Qian, Jun, Ito, Shinji, Satoh, Junko, Geng, Hongmin, Tanaka, Keisuke, Hattori, Shunji, Kojima, Kenji, Takita, Teisuke, and Yasukawa, Kiyoshi
- Subjects
COLLAGEN ,PEPSIN ,MASS spectrometry - Abstract
Bovine type I collagen consists of two α1 and one α2 chains, containing the internal triple helical regions and the N- and C-terminal telopeptides. In industries, it is frequently digested with porcine pepsin to produce a triple helical collagen without the telopeptides. However, the digestion mechanism is not precisely understood. Here, we performed a mass spectrometric analysis of the pepsin digest of the N-terminal telopeptide pQLSYGYDEKSTGISVP (1–16) in the α1 chain. When purified collagen was digested, pQLSYGY (1–6) and pQLSYGYDEKSTG (1–12) were identified, while DEKSTG (7–12) was not. When the N-terminal telopeptide mimetic synthetic peptide pQLSK(MOCAc)GYDEKSTGISK(Dnp)P-NH2was digested, pQLSK(MOCAc)GYDEKSTG (1–12) and ISK(Dnp)P-NH2(13−16) were readily identified, pQLSK(MOCAc)GY (1−6) and DEKSTGISK(Dnp)P-NH2(7−16) were weakly detected, and DEKSTG (7–12) was hardly identified. These results suggest that pepsin preferentially cleaves Tyr6–Asp7 and less preferentially Gly12–Ile13. They also suggest that the former cleavage requires native collagen structure, while the latter cleavage does not. Pepsin preferentially cleaves Tyr6–Asp7 and less preferentially Gly12–Ile13. The former cleavage requires native collagen structure, while the latter cleavage does not. [ABSTRACT FROM PUBLISHER]
- Published
- 2017
- Full Text
- View/download PDF
12. Inhibition of GluN2B pathway is involved in the neuroprotective effect of silibinin on streptozotocin-induced Alzheimer's disease models.
- Author
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Liu, Panwen, Wang, Chenkang, Chen, Wenhui, Kang, Yu, Liu, Weiwei, Qiu, Zhiyue, Hayashi, Toshihiko, Mizuno, Kazunori, Hattori, Shunji, Fujisaki, Hitomi, and Ikejima, Takashi
- Abstract
Over-activation of N -methyl-D-aspartate receptors (NMDARs) is involved in sporadic Alzheimer's disease. Silibinin, a natural flavonoid gained from the seeds of Silybum marianum , exerts neuroprotective effects on sporadic AD models, but its impacts on NMDARs remain unknown. To study silibinin's regulatory effects on NMDARs pathway in sporadic AD models. MTT assay, western blotting, confocal microscopy, flow cytometry, RT-PCR, and siRNA transfection etc. were used for cellular and molecular studies. The direct interactions between silibinin and NMDAR subunits were evaluated by computational molecular docking, drug affinity responsive target stability (DARTS) assay and cellular thermal shift assay (CETSA). Y maze test, novel objects recognition test and Morris water maze test were conducted to examine the learning and memory ability of rats. An in vitro AD model was established by treating HT22 murine hippocampal neurons with streptozotocin (STZ), as evidenced by the amyloid β (Aβ) deposition and hyperphosphorylation of tau proteins. Silibinin shows protection of neurons against STZ-induced cell damage. It is noteworthy that STZ-induced cellular calcium influx is inhibited by silibinin-treatment, indicating the possible modulation of calcium channels. Studies on NMDARs, the most widely distributed calcium channel, by using molecular docking, DARTS and CESTA, reveal that the GluN2B subunit, but not GluN2A, is the potential target of silibinin. Further studies using the pharmacological agonist (NMDA) and the GluN2B-specific inhibitor (Ifenprodil) or siRNA, indicate that the protection by silibinin treatment from STZ-induced cytotoxicity is medicated through interference with GluN2B-containing NMDARs, followed by the upregulation of CaMKIIα/ BDNF/ TrkB signaling pathway and improved levels of synaptic proteins (SYP and PSD-95). The results in vivo using rats intracerebroventricularly injected with STZ (ICV-STZ), a well-established sporadic AD model, confirm that silibinin improves learning and memory ability in association with modulation of the GluN2B/CaMKIIα/ BDNF/TrkB signaling pathway. Inhibiting over-activation of GluN2B-containing NMDARs is involved in the neuroprotective effect of silibinin on STZ-induced sporadic AD models. [Display omitted] [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
13. Exposure of bovine dermal tissue to ultraviolet light under the Antarctic ozone hole.
- Author
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Takahashi, Tetsuya, Ogura, Takayuki, Tanaka, Keisuke, Hattori, Shunji, Kudoh, Sakae, and Imura, Satoshi
- Subjects
ULTRAVIOLET radiation ,DERMIS ,OZONE layer depletion ,CROSSLINKED polymers ,COLLAGEN - Abstract
Bovine dermis was exposed outdoors in the Antarctic in 2013 to study the skin damage caused by short-wavelength ultraviolet light under the ozone hole. Collagen was extracted from the exposed dermis with pepsin. The amount of solubilized collagen in the exposed dermis was only 20%–40% of that in dermis shielded from ultraviolet light. The dermis was most difficult to extract when exposed in summer, and then when exposed in spring. Differential scanning calorimetry was used to determine the melting endothermic behavior of the dermal tissue. The peak temperature was highest for the dermis exposed in summer. The exposed dermis was degraded with cyanogen bromide to determine whether cross-linking had occurred. Cross-linked peptides were detected in the dermis exposed in summer or spring, but the dermis exposed in autumn did not differ markedly from the light-shielded dermis. These data show that cross-linkages were readily formed in the collagen molecule chains in dermis exposed to ultraviolet light in summer, when solar elevation is highest and the period of sunshine is longest. A comparison of the dermis exposed in spring and that exposed in autumn showed that cross-linkages were formed more readily by ultraviolet light in spring, when the ozone hole occurred. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
14. Characterization of Angiotensin-Converting Enzyme Inhibitory Activity of X-Hyp-Gly-Type Tripeptides: Importance of Collagen-Specific Prolyl Hydroxylation
- Author
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Taga, Yuki, Hayashida, Osamu, Ashour, Ahmed, Amen, Yhiya, Kusubata, Masashi, Ogawa-Goto, Kiyoko, Shimizu, Kuniyoshi, and Hattori, Shunji
- Abstract
Hydroxyproline (Hyp) is a collagen-specific amino acid formed by post-translational hydroxylation of Pro residues. Various Hyp-containing oligopeptides are transported into the blood at high concentrations after oral ingestion of collagen hydrolysate. Here we investigated the angiotensin-converting enzyme (ACE) inhibitory activity of X-Hyp-Gly-type tripeptides. In an in vitro assay, ginger-degraded collagen hydrolysate enriched with X-Hyp-Gly-type tripeptides dose-dependently inhibited ACE and various synthetic X-Hyp-Gly-type tripeptides showed ACE-inhibitory activity. In particular, strong inhibition was observed for Leu-Hyp-Gly, Ile-Hyp-Gly, and Val-Hyp-Gly with IC50values of 5.5, 9.4, and 12.8 μM, respectively. Surprisingly, substitution of Hyp with Pro dramatically decreased inhibitory activity of X-Hyp-Gly, indicating that Hyp is important for ACE inhibition. This finding was supported by molecular docking experiments using Leu-Hyp-Gly/Leu-Pro-Gly. We further demonstrated that prolyl hydroxylation significantly enhanced resistance to enzymatic degradation by incubation with mouse plasma. The strong ACE-inhibitory activity and high stability of X-Hyp-Gly-type tripeptides highlight their potential for hypertension control.
- Published
- 2018
- Full Text
- View/download PDF
15. Expression of collagenase in Flavobacterium psychrophilum isolated from cold-water disease-affected ayu ( Plecoglossus altivelis ).
- Author
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Nakayama, Hitoshi, Tanaka, Keisuke, Teramura, Naoko, and Hattori, Shunji
- Subjects
COLLAGENASES ,FLAVOBACTERIUM ,AYU - Abstract
The collagenase activity and thefpcolgene were examined inFlavobacterium psychrophilumisolates from cold-water disease (CWD)-affected ayu,Plecoglossus altivelis. Collagenase expression was closely related to the accumulated mortality of CWD-affected ayu. RT-qPCR and bacterial challenge experiments showed thatF. psychrophilumayu isolate WA-1 expressed thefpcolgene more actively and was more virulent than ayu isolate WA-2. The amago (Oncorhynchus masou) isolate WB-1, which possesses a pseudo-fpcolgene, was not harmful to ayu. Hitherto, the well-studied metalloproteases Fpp1 and Fpp2 have been considered virulence factors. However, the most virulent isolate against ayu (WA-1) showed no Fpp activity because of a deletion mutation or an insertion of a transposon in thefppgenes. The less virulent WA-2 isolate showed only Fpp1 activity. Taken together, these results suggest that collagenolytic activity, but not Fpp activity, is related to the virulence ofF. psychrophilumisolates in CWD-affected ayu. Comparative study ofFlavobacterium psychrophilumisolates from ayu and amago suggests that the collagenase is one of the pathogenic factors for ayu. [ABSTRACT FROM PUBLISHER]
- Published
- 2016
- Full Text
- View/download PDF
16. Production of a novel wheat gluten hydrolysate containing dipeptidyl peptidase-IV inhibitory tripeptides using ginger protease
- Author
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Taga, Yuki, Hayashida, Osamu, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Abstract
Wheat gluten is a Pro-rich protein complex comprising glutenins and gliadins. Previous studies have reported that oral intake of enzymatic hydrolysates of gluten has beneficial effects, such as suppression of muscle injury and improvement of hepatitis. Here, we utilized ginger protease that preferentially cleaves peptide bonds with Pro at the P2position to produce a novel type of wheat gluten hydrolysate. Ginger protease efficiently hydrolyzed gluten, particularly under weak acidic conditions, to peptides with an average molecular weight of <600 Da. In addition, the gluten hydrolysate contained substantial amounts of tripeptides, including Gln-Pro-Gln, Gln-Pro-Gly, Gln-Pro-Phe, Leu-Pro-Gln, and Ser-Pro-Gln (e.g. 40.7 mg/g at pH 5.2). These gluten-derived tripeptides showed high inhibitory activity on dipeptidyl peptidase-IV with IC50values of 79.8, 70.9, 71.7, 56.7, and 78.9 μM, respectively, suggesting that the novel gluten hydrolysate prepared using ginger protease can be used as a functional food for patients with type 2 diabetes.
- Published
- 2017
- Full Text
- View/download PDF
17. Cyclophilin B Deficiency Causes Abnormal Dentin Collagen Matrix
- Author
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Terajima, Masahiko, Taga, Yuki, Cabral, Wayne A., Nagasawa, Masako, Sumida, Noriko, Hattori, Shunji, Marini, Joan C., and Yamauchi, Mitsuo
- Abstract
Cyclophilin B (CypB) is an endoplasmic reticulum-resident protein that regulates collagen folding, and also contributes to prolyl 3-hydroxylation (P3H) and lysine (Lys) hydroxylation of collagen. In this study, we characterized dentin type I collagen in CypB null (KO) mice, a model of recessive osteogenesis imperfecta type IX, and compared to those of wild-type (WT) and heterozygous (Het) mice. Mass spectrometric analysis demonstrated that the extent of P3H in KO collagen was significantly diminished compared to WT/Het. Lys hydroxylation in KO was significantly diminished at the helical cross-linking sites, α1/α2(I) Lys-87 and α1(I) Lys-930, leading to a significant increase in the under-hydroxylated cross-links and a decrease in fully hydroxylated cross-links. The extent of glycosylation of hydroxylysine residues was, except α1(I) Lys-87, generally higher in KO than WT/Het. Some of these molecular phenotypes were distinct from other KO tissues reported previously, indicating the dentin-specific control mechanism through CypB. Histological analysis revealed that the width of predentin was greater and irregular, and collagen fibrils were sparse and significantly smaller in KO than WT/Het. These results indicate a critical role of CypB in dentin matrix formation, suggesting a possible association between recessive osteogenesis imperfecta and dentin defects that have not been clinically detected.
- Published
- 2017
- Full Text
- View/download PDF
18. Development of immersion vaccine for bacterial cold-water disease in ayu Plecoglossus altivelis
- Author
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Nakayama, Hitoshi, Mori, Mariko, Takita, Teisuke, Yasukawa, Kiyoshi, Tanaka, Keisuke, Hattori, Shunji, Aikawa, Hideaki, Hasegawa, Osamu, Okamura, Takashi, Takegami, Kentarou, Motokawa, Shogo, Kuwahara, Masakazu, and Amano, Kenichi
- Abstract
Flavobacterium psychrophilum(F. psychrophilum) is the causative agent of bacterial cold-water disease (BCWD) that occurs in ayu Plecoglossus altivelis. Formalin-killed cell of F. psychrophilumhas long been studied as an immersion vaccine for BCWD. In this study, we explored the possibility of F. psychrophilumcollagenase (fpcol) for use as the immersion vaccine. BCWD convalescent ayu sera contained specific IgM antibodies against somatic F. psychrophilumand fpcol, meaning that fpcol is a promising antigen for the vaccine development. The recombinant fpcol was successfully expressed in Escherichia coliand Brevibacillus chosinensis(B. chosinensis). The culture supernatant of the B. chosinensiswas used as an immersion vaccine solution. The vaccinated ayu were then challenged by soaking into F. psychrophilumculture. In two experimental groups, the relative percentages of survivals were 63 and 38%, respectively, suggesting that fpcol is promising as the immersion vaccine for ayu-BCWD.
- Published
- 2017
- Full Text
- View/download PDF
19. The cleavage site preference of the porcine pepsin on the N-terminal α1 chain of bovine type I collagen: a focal analysis with mass spectrometry
- Author
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Qian, Jun, Ito, Shinji, Satoh, Junko, Geng, Hongmin, Tanaka, Keisuke, Hattori, Shunji, Kojima, Kenji, Takita, Teisuke, and Yasukawa, Kiyoshi
- Abstract
Bovine type I collagen consists of two α1 and one α2 chains, containing the internal triple helical regions and the N- and C-terminal telopeptides. In industries, it is frequently digested with porcine pepsin to produce a triple helical collagen without the telopeptides. However, the digestion mechanism is not precisely understood. Here, we performed a mass spectrometric analysis of the pepsin digest of the N-terminal telopeptide pQLSYGYDEKSTGISVP (1–16) in the α1 chain. When purified collagen was digested, pQLSYGY (1–6) and pQLSYGYDEKSTG (1–12) were identified, while DEKSTG (7–12) was not. When the N-terminal telopeptide mimetic synthetic peptide pQLSK(MOCAc)GYDEKSTGISK(Dnp)P-NH2was digested, pQLSK(MOCAc)GYDEKSTG (1–12) and ISK(Dnp)P-NH2(13−16) were readily identified, pQLSK(MOCAc)GY (1−6) and DEKSTGISK(Dnp)P-NH2(7−16) were weakly detected, and DEKSTG (7–12) was hardly identified. These results suggest that pepsin preferentially cleaves Tyr6–Asp7 and less preferentially Gly12–Ile13. They also suggest that the former cleavage requires native collagen structure, while the latter cleavage does not.
- Published
- 2017
- Full Text
- View/download PDF
20. Synergistic repellent activity of hot and cool thermal-sense compounds against two stored product insect pests.
- Author
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Shimomura, Kenji, Suzuki, Nobuhiro, and Hattori, Shunji
- Abstract
The repellent activities of one hot thermal-sense compound, vanillyl butyl ether (VBE), and two cool thermal-sense compounds, menthoxypropanediol (MPD) and (−)-isopulegol (IPG), against two stored product insects, Tribolium confusum and Liposcelis bostrychophila , were investigated. Such thermal-sense compounds affect mammals through transient receptor potential (TRP) channels, and both types of agonists (VBE and menthol) exhibited mutually inhibitory effects on mammalian TRPV1 and TRPM8 channels. Each compound exhibited repellent activity against both T . confusum and L . bostrychophila in a dose-dependent manner. Furthermore, synergistic effects were observed between VBE and MPD or IPG. The composition of TRP channels is different between mammals and insects. These results suggest that the use of thermal-sense compounds might serve as an alternative to essential oils in stored products protection. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
21. Expression of collagenase in Flavobacterium psychrophilumisolated from cold-water disease-affected ayu (Plecoglossus altivelis)
- Author
-
Nakayama, Hitoshi, Tanaka, Keisuke, Teramura, Naoko, and Hattori, Shunji
- Abstract
The collagenase activity and the fpcolgene were examined in Flavobacterium psychrophilumisolates from cold-water disease (CWD)-affected ayu, Plecoglossus altivelis. Collagenase expression was closely related to the accumulated mortality of CWD-affected ayu. RT-qPCR and bacterial challenge experiments showed that F. psychrophilumayu isolate WA-1 expressed the fpcolgene more actively and was more virulent than ayu isolate WA-2. The amago (Oncorhynchus masou) isolate WB-1, which possesses a pseudo-fpcolgene, was not harmful to ayu. Hitherto, the well-studied metalloproteases Fpp1 and Fpp2 have been considered virulence factors. However, the most virulent isolate against ayu (WA-1) showed no Fpp activity because of a deletion mutation or an insertion of a transposon in the fppgenes. The less virulent WA-2 isolate showed only Fpp1 activity. Taken together, these results suggest that collagenolytic activity, but not Fpp activity, is related to the virulence of F. psychrophilumisolates in CWD-affected ayu.
- Published
- 2016
- Full Text
- View/download PDF
22. Site-specific Quantitative Analysis of Overglycosylationof Collagen in Osteogenesis Imperfecta Using Hydrazide Chemistry andSILAC.
- Author
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Taga, Yuki, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Published
- 2013
- Full Text
- View/download PDF
23. Protection Effect for Collagen Artificial Skin of UV-cut Materials in Antarctica.
- Author
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Takahashi, Tetsuya, Yamamoto, Tatsuyuki, Kasai, Wakako, Kondo, Tetsuo, Tanaka, Keisuke, Hattori, Shunji, Irie, Shinkichi, Kudoh, Sakae, Imura, Satoshi, and Kanda, Satoshi
- Abstract
In the Antarctica during mid-summer being appropriate for a severe environment of UV-rays, the authors surveyed the protective effect of UV-cut materials against UV-rays, using a collagen sheet (hereafter "collagen artificial skin") that was covered with a UV-cut film and subjected to exposure outdoors. First, exposure of the laminated, collagen artificial skin not covered with any film to UV-rays caused the extracts from the 1st layer, i.e. the utmost outerlayer, of the collagen artificial skin to increase the terminal amino group concentration up to as high as 3.20 nmol/ml, since the collagen molecular chain was largely cut. On the other hand, for the 2nd and 3rd layers of the said sheet, the terminal amino group concentrations were found to have decreased in order, and for the 4th layer and onward, the concentrations were shown to have decreased down to 0.22 nmol/ml, which were nearly equal to the level before the exposure. Although the bands of three molecular chains of &alpha-, β-, and γ-chains in all samples after exposure to UV-rays were disappeared, it was also shown that the more a layer was inside, the less the collagen molecular chain was not dissolved. In addition, when the collagen artificial skin covered with a UV-blocking, zinc oxide-added film was exposed to the sunlight, it was demonstrated that the more the addition of zinc oxide in the film increased, the lower the terminal amino group concentration in the extracts from the exposed, collagen artificial skin was observed. At the time when the added quantity of zinc oxide reached 0.40v%, the concentration was maintained almost at the same level as observed prior to the exposure. Also in comparison with UV-protecting agents, it was shown that the film added with an organic compound acting as a UV-absorbing agent reduced most the damages sustained by UV-rays. As described above, the evaluation method using a collagen artificial skin could be effectively employed in outdoor exposure for examining the protective effect of UV-cut materials to the skin against the UV-rays in the Antarctica. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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24. Evaluation of UV Protection Effect for UV-Cut Materials Using the Collagen Artificial Skin.
- Author
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Takahashi, Tetsuya, Tanaka, Keisuke, Hattori, Shunji, Irie, Shinkichi, Kudoh, Sakae, Imura, Satoshi, and Kanda, Satoshi
- Abstract
For preventing damages on the skin by UV-rays, various UV-cut materials have been developed. In the present study, the authors made a collagen sheet (hereafter "collagen artificial skin") to imitate the human skin on an experimental basis and, after having covered this sheet with a film added with various UV-protecting agents, subjected it artificially to UV-irradiation. Under such condition, the skin protective effect of each kind of UV-cut materials against UV rays was examined, and the various methods to evaluate damages on the sheet caused by UV rays were also reviewed. First, the UV-rays were applied to the collagen artificial skin that was not covered with the film, and the extracts were obtained from the sheet after irradiation. The analysis of the extracts by electrophoresis revealed that the bands of three molecular chains such as α-, β, and γ-chains of polypeptides were disappeared under UV-irradiation even only for 15 minutes (UV-Energy : 7.58x10
4 J/m², at 310nm). When the irradiation time reached 25 hours (UV-Energy: 7.58x106 J/m², at 310nm), hydrolysates from the collagen in the sheet started to appear, and when the irradiation time exceeded further 200 hours (UV-Energy: 6.06x107 7J/m², at 310nm), a considerable amount of hydrolysates was recognized. Also it was found that the longer the irradiation time of UV-rays, the higher the concentration of the terminal amino group in the extracts from the collagen was observed. Then, the UV-rays were irradiated to the collagen artificial skin added with various UV-scattering agents. As a result, addition of UV-scattering agents to the film of any of the materials reduced scission of the collagen molecular chains caused by UV-rays, resulting in reduction of the increase of the terminal amino group concentration. From the above results, it was suggested that the use of a collagen artificial skin was one of the effective measures as a method for examining the protective effect against UV-rays. [ABSTRACT FROM AUTHOR]- Published
- 2009
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- View/download PDF
25. Collagen as a Animal Derived Fibrous Protein-its Character and Application.
- Author
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HATTORI, SHUNJI
- Published
- 2009
- Full Text
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26. Improved Physical and Biochemical Features of a Collagen Membrane by Conjugating with Soluble Egg Shell Membrane Protein.
- Author
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Ino, Tarnao, Hattori, Makoto, Yoshida, Tadashi, Hattori, Shunji, Yoshimura, Keiji, and Takahashii, Koji
- Subjects
CELL membranes ,COLLAGEN ,PEPSIN ,MEMBRANE proteins ,FIBROBLASTS ,KERATINOCYTES - Abstract
The article discusses the results of a study which examined the improvement in the physical and biochemical characteristics of a pepsin-solubilized collagen (PSC) membrane by conjugating with soluble egg shell membrane protein (SEP). The conjugated membrane was found to have thicker collagen fibrils and higher stability than PSC membrane alone. An SEP-PSC coating accelerated the growth of fibroblasts. However, the cell growth of keratinocytes did not change.
- Published
- 2006
- Full Text
- View/download PDF
27. Epigallocatechin-3-O-gallate Inhibits Fibroblast Contraction of Floating Collagen Gel: Interaction between Epigallocatechin-3-O-gallate and Platelet Derived Growth Factor.
- Author
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Suzuki, Yasuo, Hattori, Shunji, and Isemura, Mamoru
- Subjects
CATECHIN ,COLLAGEN ,COLLOIDS ,FIBROBLASTS ,PLATELET-derived growth factor ,ENZYME-linked immunosorbent assay - Abstract
Investigates the effects of (-)-epigallocatechin gallate (EGCG) on the contraction of floating collagen gel by fibroblasts. Interaction between EGCG and platelet-derived growth factor; Affinity chromatography; Enzyme-linked immunoassay; Mechanism of inhibition of collagen gel contraction by EGCG.
- Published
- 2004
- Full Text
- View/download PDF
28. Stable Isotope-Labeled Collagen: A Novel and Versatile Tool for Quantitative Collagen Analyses Using Mass Spectrometry
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Taga, Yuki, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Abstract
Collagens are the most abundant proteins in animals and are involved in many physiological/pathological events. Although various methods have been used to quantify collagen and its post-translational modifications (PTMs) over the years, it is still difficult to accurately quantify type-specific collagen and minor collagen PTMs. We report a novel quantitative method targeting collagen using stable isotope-labeled collagen named “SI-collagen”, which was labeled with isotopically heavy lysine, arginine, and proline in fibroblasts culture. We prepared highly labeled and purified SI-collagen for use as an internal standard in mass spectrometric analysis, particularly for a new approach using amino acid hydrolysis. Our method enabled accurate collagen analyses, including quantification of (1) type-specific collagen (types I and III in this paper), (2) total collagen, and (3) collagen PTMs by LC–MS with high sensitivity. SI-collagen is also applicable to other diverse analyses of collagen and can be a powerful tool for various studies, such as detailed investigation of collagen-related disorders.
- Published
- 2014
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- View/download PDF
29. Site-specific Quantitative Analysis of Overglycosylation of Collagen in Osteogenesis Imperfecta Using Hydrazide Chemistry and SILAC
- Author
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Taga, Yuki, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Abstract
We recently developed a novel method for analysis of collagen O-glycosylations, which include galactosyl-hydroxylysine (GHL) and glucosyl-galactosyl-hydroxylysine (GGHL), using hydrazide chemistry (Taga, Y., Mol. Cell. Proteomics2012, 11(6), M111.010397). Here we investigated an overglycosylation model of collagen produced by cultured skin fibroblasts from osteogenesis imperfecta (OI) patients using this method. Many GHL/GGHL sites were identified in normal and OI type I collagens by LC–MS analysis after the glycopeptide purification procedure. Further, relative quantification was performed on each identified glycopeptide using stable isotope labeling by amino acids in cell culture (SILAC). Significant increases of GGHL were observed at respective glycosylation sites of type I collagen in OI, whereas an OI-specific glycosylation site was not found. These results demonstrated that the overglycosylation of type I collagen proceeds only at specific sites, resulting in accumulation of GGHL, rather than because of an increase of nonspecific glycosylation. Although the roles of collagen O-glycosylations in OI and even in normal conditions are still incompletely understood, the location of GHL/GGHL in the collagen sequence is suggested to be important for their functions.
- Published
- 2013
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- View/download PDF
30. Bi-phasic effect of gelatin in myogenesis and skeletal muscle regeneration
- Author
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Liu, Xiaoling, Zu, Er, Chang, Xinyu, Ma, Xiaowei, Wang, Ziqi, Song, Xintong, Li, Xiangru, Yu, Qing, Kamei, Ken-ichiro, Hayashi, Toshihiko, Mizuno, Kazunori, Hattori, Shunji, Fujisaki, Hitomi, Ikejima, Takashi, and Wang, Dan Ohtan
- Abstract
Skeletal muscle regeneration requires extracellular matrix (ECM) remodeling, including an acute and transient breakdown of collagen that produces gelatin. Although the physiological function of this process is unclear, it has inspired the application of gelatin to injured skeletal muscle for a potential pro-regenerative effect. Here, we investigated a bi-phasic effect of gelatin in skeletal muscle regeneration, mediated by the hormetic effects of reactive oxygen species (ROS). Low-dose gelatin stimulated ROS production from NADPH oxidase 2 (NOX2) and simultaneously upregulated the antioxidant system for cellular defense, reminiscent of the adaptive compensatory process during mild stress. This response triggered the release of the myokine IL-6, which stimulates myogenesis and facilitates muscle regeneration. By contrast, high-dose gelatin stimulated ROS overproduction from NOX2 and the mitochondrial chain complex, and ROS accumulation by suppressing the antioxidant system, triggering the release of TNFα, which inhibits myogenesis and regeneration. Our results have revealed a bi-phasic role of gelatin in regulating skeletal muscle repair mediated by intracellular ROS, the antioxidant system and cytokine (IL-6 and TNFα) signaling.
- Published
- 2021
- Full Text
- View/download PDF
31. Improved Physical and Biochemical Features of a Collagen Membrane by Conjugating with Soluble Egg Shell Membrane Protein
- Author
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INO, Tamao, HATTORI, Makoto, YOSHIDA, Tadashi, HATTORI, Shunji, YOSHIMURA, Keiji, and TAKAHASHI, Koji
- Abstract
Soluble egg shell membrane protein (SEP) prepared from the egg shell membrane by performic acid oxidation and pepsin digestion was applied to prepare a membrane conjugated with pepsin-solubilized collagen (PSC) by cross-linking with water-soluble carbodiimide. SEP enabled a conjugated membrane to be constructed with a well-developed network structure having thicker collagen fibrils and higher stability than the PSC membrane alone, as indicated by reduced solubility and collagenase-digestion, increased denaturation temperature, and superior flexibility. The SEP-PSC membrane (SEP:PSC of 1:1, w/w) had lower sweatiness than the PSC membrane. Fibroblasts and keratinocytes adhered to the SEP-PSC-coated wells (SEP:PSC of 1:10, w/w) as a fundamental model of an artificial skin similar to that to the PSC-coated wells due to similar expression level of several cell-adhesion proteins. An SEP-PSC coating accelerated the cell growth of fibroblasts, whereas the cell growth of keratinocytes did not change, or decreased slightly due to more rapid differentiation indicated by a more substantial expression of differentiation marker proteins than a PSC-coating without SEP.
- Published
- 2006
- Full Text
- View/download PDF
32. Epigallocatechin-3-O-gallate Inhibits Fibroblast Contraction of Floating Collagen Gel: Interaction between Epigallocatechin-3-O-gallate and Platelet Derived Growth Factor
- Author
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SUZUKI, Yasuo, HATTORI, Shunji, and ISEMURA, Mamoru
- Abstract
The effects of (−)-epigallocatechin gallate (EGCG) on the contraction of floating collagen gel by fibroblasts were investigated. EGCG inhibited collagen gel contraction dose-dependently. On the basis of the fact that platelet-derived growth factor (PDGF) is one of the serum components with stimulatory activity in collagen gel contraction, we examined the possibility that interaction between EGCG and PDGF may be involved in this inhibition mechanism. We confirmed this by recombinant PDGF-BB in the present system and we found that EGCG inhibited PDGF-stimulated collagen gel contraction. The results of affinity chromatography indicated that PDGF was bound by EGCG immobilized on agarose gel as detected by enzyme-linked immunoassay and Western blotting. These findings suggest that binding of EGCG to PDGF is at least partly involved in the mechanism of inhibition of collagen gel contraction by EGCG.
- Published
- 2004
- Full Text
- View/download PDF
33. Analysis of the major epitope of the α2 chain of bovine type I collagen in children with bovine gelatin allergy
- Author
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Hori, Hisae, Hattori, Shunji, Inouye, Sakae, Kimura, Akinori, Irie, Shinkichi, Miyazawa, Hiroshi, and Sakaguchi, Masahiro
- Abstract
Background:Anaphylaxis to measles, mumps, and rubella vaccines has been reported. It has been found that most of these reactions to live vaccines are caused by type I allergy with the bovine gelatin present in the vaccines as an allergen. Gelatin mainly includes denatured type I collagen, which consists of α1 and α2 chains. We previously reported that allergic reactions to gelatin are caused by the type I collagen α2 (α2[I]) chain. Objective:To aid in the development of gelatin that has little or no allergenicity in human subjects, we investigated epitopes of bovine α2(I) chain with use of IgE in gelatin-sensitive children. Methods:Serum samples were collected from 15 patients who had systemic allergic reactions to vaccines and high levels of specific IgE to bovine gelatin. Eleven overlapping recombinant proteins that cover bovine α2(I) were prepared with a bacterial expression vector. We examined IgE reactivity to these recombinant proteins by means of ELISA. Fifteen peptides covering a major reactive recombinant protein were synthesized. The IgE-reacting epitope was identified by means of IgE-ELISA inhibition with these synthetic peptides and pooled serum from the patients. Results:We found that of the 15 patients, 13 showed IgE reactivity to a recombinant protein (no. 3) spanning the central region of the collagenous domain (418Gly-662Pro). Furthermore, all 13 patients showed IgE reactivity to the 4-kd recombinant protein (no. 3a) spanning the region from 461Pro to 500Glu. In IgE-ELISA inhibition we found that a minimum IgE epitope of gelatin allergen was composed of the 10-amino-acid sequence 485Ile-Pro-Gly-Glu-Phe-Gly-Leu-Pro-Gly-Pro494. This sequence is not observed in the human type I collagen α1 and α2 chains, nor is it found in the bovine type I collagen α1 chain. Conclusions:We found that Ile-Pro-Gly-Glu-Phe-Gly-Leu-Pro-Gly-Pro is a major IgE epitope of the α2 chain of bovine type I collagen in patients with gelatin allergy. The degree of anaphylaxis to gelatin in vaccines might be reduced by digestion of this IgE-binding site in gelatin. (J Allergy Clin Immunol 2002;110:652-7.)
- Published
- 2002
- Full Text
- View/download PDF
34. Systematic Separation and Purification of Elastase, Gelatinase (Matrix Metalloproteinase 9), and Collagenase (Matrix Metalloproteinase 8) from Polymorphonuclear Leukocytes in Dialyzers Previously Used by Patients with Renal Failure
- Author
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Kobayashi, Takashi, Nishikawa, Takeji, Hattori, Shunji, Yoshida, Naoki, Takagi, Tamotsu, Watanabe, Hideto, Hori, Hisae, and Nagai, Yutaka
- Abstract
We developed a simple and effective method for the systematic separation and purification of human polymorphonuclear leukocyte (PMN) proteinases, elastase, gelatinase (matrix metalloproteinase 9, type IV collagenase), and collagenase (matrix metalloproteinase 8), derived from the extracts of hollow fiber dialyzers that had been utilized in the treatment of patients with renal failure. The fraction containing elastase was grossly separated from that containing gelatinase and collagenase by heparin-Sepharose chromatography and purified in an aprotinin column. The remaining two enzymes were then separated using the gelatin-Sepharose column after gel chromatography following ammonium sulfate precipitation. Gelatinase and collagenase were further purified by gelatin-Sepharose chromatography as a latent form and by collagen-Sepharose chromatography as an activated form. This novel method offers procedural advantages over existing methods that separate PMNs from the whole blood of volunteers for experimental research purposes.
- Published
- 2001
- Full Text
- View/download PDF
35. Effect of IL-12 Encoding Plasmid Administration on Tight-Skin Mouse
- Author
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Tsuji-Yamada, Junko, Nakazawa, Masatoshi, Takahashi, Kazuo, Iijima, Katsumasa, Hattori, Shunji, Okuda, Kenji, Minami, Mutsuhiko, Ikezawa, Zenro, and Sasaki, Tetsuo
- Abstract
The tight-skin (Tsk/+) mutant mice, a putative murine model of scleroderma, are characterized by the excessive deposition of collagen and the presence of antinuclear antibodies. Type 2 cytokines, such as IL-4 and IL-6, are capable of regulating the synthesis of various matrix molecules, including type I collagen, by fibroblasts. IL-12 is well known to induce type 1 cytokine production and to reduce type 2 activity. Here, we examined the effect of IL-12 encoding plasmid (pCAGGSIL-12) on the disease progression of Tsk/+ mice. pCAGGSIL-12 plasmid or pCAGGS parental vector was injected intramuscularly 7 times at 3 week intervals into Tsk/+ mice. One week after the last injection, pCAGGSIL-12 administered Tsk/+ mice exhibited a marked decrease in the skin thickness compared with the mice treated with pCAGGS vector. The serum levels of antinuclear antibodies were diminished in pCAGGSIL-12 treated mice. IL-4 production by spleen cells from pCAGGSIL-12 plasmid treated mice was significantly lower than that from vector treated mice. These results indicate that pCAGGSIL-12 administration into Tsk/+ mice had beneficial effects in preventing the collagen accumulation in the skin and suppressing the autoimmunity via improvement of Th1/Th2 balance. The present study suggests that the IL-12 encoding plasmid administration might have a therapeutic effect on systemic sclerosis.
- Published
- 2001
- Full Text
- View/download PDF
36. Possible Involvement of Aminotelopeptide in Self-assembly and Thermal Stability of Collagen I as Revealed by Its Removal with Proteases*
- Author
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Sato, Kaori, Ebihara, Tetsuya, Adachi, Eijiro, Kawashima, Seiichi, Hattori, Shunji, and Irie, Shinkichi
- Abstract
The functions of aminotelopeptide and N-terminal cross-linking of collagen I were examined. Acetic acid-soluble collagen I (ASC) was purified from neonatal bovine skin and treated with three kinds of proteases. The amino acid sequencing analysis of the N terminus showed that ASC contained a full-length aminotelopeptide. Pepsin and papain cleaved the aminotelopeptide of the α1 chain at the same site and the aminotelopeptide of the α2 chain at different sites. Proctase-treated ASC lost the whole aminotelopeptide, and the N-terminal sequence began from the tenth residue inside the triple helical region. The rates of fibril formation of pepsin-treated ASC and proctase-treated ASC were the same and were slower than that of ASC. The denaturation temperatures, monitored by CD ellipticity at 221 nm, of ASC, pepsin-treated, or papain-treated collagens were the same at 41.8 °C. Proctase-treated ASC showed a lower denaturation temperature of 39.9 °C. We also observed the morphology of the collagen fibrils under an electron microscope. The ASC fibrils were straight and thin, whereas the fibrils of pepsin-treated ASC were slightly twisted, and the fibrils from papain- and proctase-treated ASC were highly twisted and thick. When the collagen gel strength was examined by a modified method of viscosity-measurement, ASC was the strongest, followed by pepsin-treated ASC, and papain- and proctase-treated ASCs were the weakest. These results suggest that the aminotelopeptide plays important roles in fibril formation and thermal stability. In addition, the functions of intermolecular cross-linking in aminotelopeptides may contribute to the formation of fibrils in the correct staggered pattern and to strengthening the collagen gel.
- Published
- 2000
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37. Laminin 5 Promotes Activation and Apoptosis of the T Cells Expressing α3β1 Integrin
- Author
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Sato, Kaori, Katagiri, Koko, Hattori, Shunji, Tsuji, Tsutomu, Irimura, Tatsurou, Irie, Shinkichi, and Katagiri, Takuya
- Abstract
By introducing an α3 gene-containing plasmid into a human T cell line Jurkat, we prepared the T cells, which express a high level of the α3β1 integrin, to assess the role of laminin 5 in the skin immune system. The α3β1-expressing T cells adhered to laminin 5 and exhibited spreading. These adhered T cells showed a significant tyrosine phosphorylation of intracellular proteins including p59fynupon T-cell receptor (TCR) stimulation. Six hours after cross-linking TCR, these cells on laminin 5 secreted a three times higher level of IL-2 than those on a BSA-coated plate. Twenty hours after the stimulation, 48% of the α3β1-expressing T cells on laminin 5 caused apoptosis. The protein level of cyclin D3 and E decreased, while that of p53 increased in these T cells. These data suggest that laminin 5 may play at least two regulatory roles for T cell functions: augmentation of IL-2 production by antigen-stimulated T cells and induction of apoptosis in these T cells.
- Published
- 1999
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38. A Case of Linear IgA Disease: An Immunofluorescent Study Using Confocal Laser Scan Microscopy
- Author
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Kobayashi, Takashi, Konohana, Akira, and Hattori, Shunji
- Abstract
A 79‐year‐old Japanese woman presented with erythema and bullae on her trunk and limbs. Histological examination of the skin lesions showed subepidermal bullae and polymorphonuclear leukocyte infiltration into the papillary dermis. A direct immunofluorescent study showed the linear deposition of IgA, but not of IgG or IgM, in the basement membrane zone. Indirect immunofluorescence of the serum using confocal laser scan microscopy showed IgA, but not IgG, reactivity in the basement membrane zone. In double immunostaining experiments, IgA reactivity was also observed on the epidermal side; laminin 5 was detected on the dermal side.
- Published
- 1997
- Full Text
- View/download PDF
39. Secretion of Different Types of Gelatinases from Cultured Human Keratinocytes
- Author
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Kobayashi, Takashi, Hattori, Shunji, Nagai, Yutaka, Sakuraoka, Koichi, and Nishikawa, Takeji
- Abstract
We investigated gelatinolytic enzymes derived from cultured human keratinocytes. Using a zymograph, we detected 92 kDa and 72 kDa gelatinases as major components and 83 kDa as a minor component from a conditioned cultured medium (BSL‐K110, Kyokutoseiyaku Co.) of keratinocytes. After several passages, the 72 kDa band disappeared, and the 83 kDa band became dominant in another conditioned culture medium (K‐GM, Kurashikibouseki Co.); these keratinocytes had a relatively differentiated appearance. These results suggest that some mechanism may regulate the secretion of selected types of gelatinase from keratinocytes under different conditions.
- Published
- 1997
- Full Text
- View/download PDF
40. The complete cDNA coding sequence for the bovine Proα2(I) chain of type I procollagen
- Author
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Shirai, Tomoko, Hattori, Shunji, Sakaguchi, Masahiro, Inouye, Sakae, Kimura, Akinori, Ebihara, Tetsuya, Irie, Shinkichi, Nagai, Yutaka, and Hori, Hisae
- Abstract
The complete sequence of the cDNA for the proα2(I) chain of bovine type I procollagen is presented. The encoded amino acid sequence shows 92.0% identity to the human proα2(I) collagen chain.
- Published
- 1998
- Full Text
- View/download PDF
41. The complete cDNA coding sequence for the bovine Proα2(I) chain of type I procollagen
- Author
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Shirai, Tomoko, Hattori, Shunji, Sakaguchi, Masahiro, Inouye, Sakae, Kimura, Akinori, Ebihara, Tetsuya, Irie, Shinkichi, Nagai, Yutaka, and Hori, Hisae
- Abstract
The complete sequence of the cDNA for the proα2(I) chain of bovine type I procollagen is presented. The encoded amino acid sequence shows 92.0% identity to the human proα2(I) collagen chain.
- Published
- 1998
- Full Text
- View/download PDF
42. In-depth correlation analysis demonstrates that 4-hydroxyproline at the Yaa position of Gly-Xaa-Yaa repeats dominantly stabilizes collagen triple helix
- Author
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Taga, Yuki, Tanaka, Keisuke, Hattori, Shunji, and Mizuno, Kazunori
- Abstract
•4Hyp at the Yaa position of Gly-Xaa-Yaa repeats has the highest correlation with collagen denaturation temperature (Td), especially in vertebrates.•Significant correlation with Tdexists for Gly-Xaa-4Hyp tripeptides, but not for Gly-Pro-Yaa tripeptides.•The in-depth correlation analysis demonstrates the dominating role of Yaa position 4Hyp for collagen stability.
- Published
- 2021
- Full Text
- View/download PDF
43. Double Immunodetection of Proteins Transferred onto a Membrane Using Two Different Chemiluminescent Reagents
- Author
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Hattori, Shunji and Fujisaki, Hitomi
- Published
- 1996
- Full Text
- View/download PDF
44. Development of a Novel Method for Analyzing Collagen O-glycosylations by Hydrazide Chemistry
- Author
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Taga, Yuki, Kusubata, Masashi, Ogawa-Goto, Kiyoko, and Hattori, Shunji
- Abstract
In recent years, glycopeptide purification by hydrazide chemistry has become popular in structural studies of glycoconjugates; however, applications of this method have been almost completely restricted to analysis of the N-glycoproteome. Here we report a novel method for analyzing O-glycosylations unique to collagen, which are attached to hydroxylysine and include galactosyl-hydroxylysine and glucosyl-galactosyl-hydroxylysine. We established a hydrazide chemistry-based glycopeptide purification method using (1) galactose oxidase to introduce an aldehyde into glycopeptides and (2) formic acid with heating to elute the bound glycopeptides by cleaving the hydrazone bond. This method allows not only identification of O-glycosylation sites in collagen but also concurrent discrimination of two types of carbohydrate substitutions. In bovine type I and type II collagens, galactosyl-hydroxylysine /glucosyl-galactosyl-hydroxylysine -containing peptides were specifically detected on subsequent comprehensive liquid chromatography (LC)/MS analysis, and many O-glycosylation sites, including unreported ones, were identified. The position of glycosylated hydroxylysine, which is determined by our unambiguous and simple method, could provide insight into the physiological role of the modifications.
- Published
- 2012
- Full Text
- View/download PDF
45. Specific function of the multimeric fibronectin formed in vitro as revealed by the interaction with HT-1080 cells
- Author
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Sakai, Keiko, Fujisaka, Hitomi, Adachi, Eijiro, Hattori, Shunji, and Hayashi, Toshihiko
- Published
- 1996
- Full Text
- View/download PDF
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