24 results on '"Fang, Caiyun"'
Search Results
2. Global Analysis of Endogenously Intact S‑Acylated Peptides Reveals Localization Differentiation of Heterogeneous Lipid Chains in Mammalian Cells.
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Ji, Guanghui, Wu, Roujun, Zhang, Lei, Yao, Jun, Zhang, Cheng, Zhang, Xiaoqin, Liu, Zhiyong, Liu, Yang, Wang, Ting, Fang, Caiyun, and Lu, Haojie
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- 2023
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3. Global Analysis of Endogenously Intact S-Acylated Peptides Reveals Localization Differentiation of Heterogeneous Lipid Chains in Mammalian Cells
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Ji, Guanghui, Wu, Roujun, Zhang, Lei, Yao, Jun, Zhang, Cheng, Zhang, Xiaoqin, Liu, Zhiyong, Liu, Yang, Wang, Ting, Fang, Caiyun, and Lu, Haojie
- Abstract
S-acylation is a widespread lipidation form in eukaryotes in which various fatty acids can be covalently attached to specific cysteine residues. However, due to the low reactivity of the lipid moieties and lack of specific antibodies, purification of intact S-acylated peptides remains challenging. Here, we developed a pretreatment method for direct separation and global analysis of endogenously intact S-acylated peptides by nanographite fluoride-based solid-phase extraction (nGF-SPE), together with the investigation and optimization of the enrichment procedure as well as the LC-MS/MS analysis process. Consequently, we performed the first global profiling of endogenously intact S-acylated peptides, with 701 S-palmitoylated peptides from HeLa cell lysates in a restricted search. Furthermore, coupling the nGF-SPE method with open search mode, altogether 1119 intact S-acylated peptides were identified with the attached palmitate, palmitoleate, myristate, and octanoate chain, respectively, providing a global insight into the endogenously heterogeneous modification state. Notably, we found and validated that S-palmitoleoylation (C16:1) provided less affinity toward lipid rafts compared with S-palmitoylation (C16:0). This study developed the first straightforward way to characterize endogenously intact S-acylated peptides on a proteome-wide scale, providing the modified residues together with their attached lipid moieties simultaneously, which paves the way for further understanding of protein S-acylation.
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- 2023
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4. Differentiation of Lung Metastases Originated From Different Primary Tumors Using Radiomics Features Based on CT Imaging.
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Shang, Hui, Li, Jizhen, Jiao, Tianyu, Fang, Caiyun, Li, Kejian, Yin, Di, and Zeng, Qingshi
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Rationale and Objectives: To explore the feasibility of differentiating three predominant metastatic tumor types using lung computed tomography (CT) radiomics features based on supervised machine learning.Materials and Methods: This retrospective analysis included 252 lung metastases (LM) (from 78 patients), which were divided into the training (n = 176) and test (n = 76) cohort randomly. The metastases originated from colorectal cancer (n = 97), breast cancer (n = 87), and renal carcinoma (n = 68). An additional 77 LM (from 35 patients) were used for external validation. All radiomics features were extracted from lung CT using an open-source software called 3D slicer. The least absolute shrinkage and selection operator (LASSO) method selected the optimal radiomics features to build the model. Random forest and support vector machine (SVM) were selected to build three-class and two-class models. The performance of the classification model was evaluated with the area under the receiver operating characteristic curve (AUC) by two strategies: one-versus-rest and one-versus-one.Results: Eight hundred and fifty-one quantitative radiomics features were extracted from lung CT. By LASSO, 23 optimal features were extracted in three-class, and 25, 29, and 35 features in two-class for differentiating every two of three LM (colorectal cancer vs. renal carcinoma, colorectal cancer vs. breast cancer, and breast cancer vs. renal carcinoma, respectively). The AUCs of the three-class model were 0.83 for colorectal cancer, 0.79 for breast cancer, and 0.91 for renal carcinoma in the test cohort. In the external validation cohort, the AUCs were 0.77, 0.83, and 0.81, respectively. Swarmplot shows the distribution of radiomics features among three different LM types. In the two-class model, high accuracy and AUC were obtained by SVM. The AUC of discriminating colorectal cancer LM from renal carcinoma LM was 0.84, and breast cancer LM from colorectal cancer LM and renal carcinoma LM were 0.80 and 0.94, respectively. The AUCs were 0.77, 0.78, and 0.84 in the external validation cohort.Conclusion: Quantitative radiomics features based on Lung CT exhibited good discriminative performance in LM of primary colorectal cancer, breast cancer, and renal carcinoma. [ABSTRACT FROM AUTHOR]- Published
- 2023
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5. Specific and Reversible Enrichment of Early-Stage Glycated Proteome Based on Thiazolidine Chemistry and Palladium-Mediated Cleavage.
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Wu, Linlin, Fei, Weiwei, Liu, Zhiyong, Zhang, Lei, Fang, Caiyun, and Lu, Haojie
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- 2022
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6. FluoroTRAQ: Quantitative Analysis of Protein S‑Nitrosylation through Fluorous Solid-Phase Extraction Combining with iTRAQ by Mass Spectrometry.
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Zhang, Cheng, Xu, Yaoyao, Wang, Guoli, Fang, Caiyun, Bao, Huimin, Zhang, Ying, and Lu, Haojie
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- 2020
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7. Site-Specific Quantification of Protein Ubiquitination on MS2 Fragment Ion Level via Isobaric Peptide Labeling
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Cao, Ting, Zhang, Lei, Zhang, Ying, Yan, Guoquan, Fang, Caiyun, Bao, Huimin, and Lu, Haojie
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Proteome-wide quantitative analysis of protein ubiquitination is important to gain insight into its various cellular functions. However, it is still challenging to monitor how ubiquitination at each individual lysine residue is independently regulated, especially the whereabouts of peptides containing more than one ubiquitination site. In recent years, isobaric peptide termini labeling has been considered a promising strategy in quantitative proteomics, benefiting from its high accuracy by quantifying with a series of b, y fragment ion pairs. Herein, we extended the concept of isobaric peptide termini labeling to large-scale quantitative analysis of protein ubiquitination. A novel MS2 fragment ion based quantitative approach was developed, allowing the quantification of ubiquitination at site level via isobaric K-ε-GG peptide labeling, which combined metabolic labeling, K-ε-GG immunoaffinity enrichment, and site-selective N-terminus dimethylation. The feasibility of this proposed strategy was demonstrated through the ubiquitin proteome analysis of differently labeled MCF-7 cell digests. As a result, 2970 unique K-ε-GG peptides of 1383 proteins containing 2874 ubiquitinated sites were confidently quantified with high accuracy and sensitivity. In addition, we demonstrated that quantification on MS2 fragment ion level makes it possible to precisely quantify each individual ubiquitinated lysine residue in 39 K-ε-GG peptides bearing two ubiquitination sites by the use of specific ubiquitinated b, y ion pairs. It is expected that this proposed approach will serve as a powerful tool to quantify ubiquitination at the site level, especially for those multiubiquitinated peptides.
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- 2024
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8. Integrated Strategy for Discovery and Validation of Glycated Candidate Biomarkers for Hemodialysis Patients with Cardiovascular Complications
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Wu, Linlin, Fang, Caiyun, Zhang, Lei, Yuan, Wenjuan, Yu, Xiaofang, and Lu, Haojie
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Glycation plays a pathogenic role in many age-related degenerative pathological conditions, such as diabetes, end-stage renal diseases, and cardiovascular diseases. Mass spectrometry-based qualitative and quantitative analysis methods have been greatly developed and contribute to our understanding of protein glycation. However, it is still challenging to sensitively and accurately quantify endogenous glycated proteome in biological samples. Herein, we proposed an integrated and robust quantitative strategy for comprehensive profiling of early-stage glycated proteome. In this strategy, a filter-assisted sample preparation method was applied to reduce sample loss and improve reproducibility of sample preparation, contributing to high-throughput analysis and accurate quantification of endogenous glycated proteins with low abundance. Standard glycated peptides were spiked and performed the subsequent process together with complex samples both in label-free quantification and multiple reaction monitoring (MRM) analysis, contributing to the improvement of quantitative accuracy. In parallel, a novel approach was developed for the synthesis of heavy isotope-labeled glycated peptides used in MRM analysis. By this way, a total of 1128 endogenous glycated peptides corresponding to 203 serum proteins were identified from 60 runs of 10 pairs of hemodialysis patients with and without cardiovascular complications, and 234 glycated peptides corresponding to 63 proteins existed in >70% runs, among which 17 peptides were discovered to be differentially glycated (P< 0.05, fold-change > 1.5 or <0.67). Furthermore, we validated the glycation difference of four target peptides in 46 serum samples using MRM analysis, which were consistent with our results of label-free quantification.
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- 2021
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9. FluoroTRAQ: Quantitative Analysis of Protein S-Nitrosylation through Fluorous Solid-Phase Extraction Combining with iTRAQ by Mass Spectrometry
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Zhang, Cheng, Xu, Yaoyao, Wang, Guoli, Fang, Caiyun, Bao, Huimin, Zhang, Ying, and Lu, Haojie
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S-Nitrosylation is an important post-translational modification that occurs on cysteine amino acid and regulates signal transduction in diverse cell processes. Dysregulation of protein nitrosylation has shown close association with cardiovascular and neurological diseases, thus demanding further precise and in-depth understanding. Mass spectrometry-based proteomics has been the method of choice for analyzing S-nitrosylated (SNO-) proteins. However, due to their extremely low expression level and rapid turnover rate, quantitative analysis of the S-nitrosylation at the proteomic level remains challenging. Herein, we developed a novel approach termed FluoroTRAQ, which combined the fluorous solid-phase extraction of SNO-peptides and iTRAQ labeling for the quantitative analysis of the SNO-proteome with high sensitivity and specificity. This new analytical strategy was subsequently applied to examine the dynamic SNO-proteome changes of human umbilical vein endothelial cells upon in vitro S-nitrosoglutathione induction. Our data identified a number of novel SNO-proteins and revealed their temporal modulation as validated by biotin switch assay. Our study offered a practical approach for quantitative analysis of protein S-nitrosylation.
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- 2020
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10. Rapid and Easy Enrichment Strategy for Naturally Acetylated N Termini Based on LysN Digestion and Amine-Reactive Resin Capture
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Du, Xiaoxian, Lu, Jingtian, Yang, Lijun, Bao, Huimin, Zhang, Lei, Yan, Guoquan, Fang, Caiyun, and Lu, Haojie
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Protein N-terminal acetylation (Nα-acetylation) is one of the most common modifications in both eukaryotes and prokaryotes. Although studies have shown that Nα-acetylation plays important roles in protein assembly, stability, and location, the physiological role has not been fully elucidated. Therefore, a robust and large-scale analytical method is important for a better understanding of Nα-acetylation. Here, an enrichment strategy was presented based on LysN digestion and amine-reactive resin capture to study naturally acetylated protein N termini. Since LysN protease cleaves at the amino-terminus of the lysine residue, all resulting peptides except naturally acetylated N-terminal peptides contain free amino groups and can be removed by coupling with AminoLink Resin. Therefore, the naturally acetylated N-terminal peptides were left in solution and enriched for further liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. The method was very simple and fast, which contained no additional chemical derivatization except protein reduction and alkylation necessarily needed in bottom-up proteomics. It could be used to study acetylated N termini from complex biological samples without bias toward different peptides with various physicochemical properties. The enrichment specificity was above 99% when it was applied in HeLa cell lysates. Neo-N termini generated by endogenous degradation could be directly distinguished without the use of stable-isotope labeling because no chemical derivatization was introduced in this method. Furthermore, this method was highly complementary to the traditional analytical methods for protein N termini based on trypsin only with ArgC-like activity. Therefore, the described method was beneficial to naturally acetylated protein N termini profiling.
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- 2020
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11. In-Depth Analysis of C Terminomes Based on LysC Digestion and Site-Selective Dimethylation.
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Du, Xiaoxian, Fang, Caiyun, Yang, Lijun, Bao, Huimin, Zhang, Lei, Yan, Guoquan, and Lu, Haojie
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- 2019
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12. In-Depth Analysis of C Terminomes Based on LysC Digestion and Site-Selective Dimethylation
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Du, Xiaoxian, Fang, Caiyun, Yang, Lijun, Bao, Huimin, Zhang, Lei, Yan, Guoquan, and Lu, Haojie
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Analysis of protein C termini is very important for functional annotations of proteomes, while proteome-wide C termini analysis still poses substantial challenges. Here we described a simple and robust strategy for specific isolation of protein C termini based on LysC digestion and site-selective dimethylation to deplete N-terminal and internal peptides by scavenger materials. The performance of LysC digestion and conditions of site-selective dimethylation and resin coupling were discussed in detail. Then the strategy was successfully applied to the characterization of protein C termini of HeLa cells. A total of 781 protein C termini were identified with a 300 μg digest in our study, among which 38.9% were actually not identifiable using current trypsin digestion-based methods due to their inappropriate peptide length for MS analysis, indicating that our method was highly complementary to the existing methods. The enrichment procedure was rapid and easy to operate and could afford a very good identification efficiency by obtaining the largest C termini data set of the human proteome with the least sample loading. This method was without bias toward physicochemical properties of peptides. Moreover, a peptide-centric database was first introduced to analyze protein C termini, which effectively improved the accuracy and speed of the database search. Therefore, our method can be used to effectively and selectively isolate protein C termini and contributes to the global annotation of C terminomes.
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- 2019
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13. Selective Identification and Site-Specific Quantification of 4-Hydroxy-2-nonenal-Modified Proteins
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Zhang, Siwen, Fang, Caiyun, Yuan, Wenjuan, Zhang, Ying, Yan, Guoquan, Zhang, Lei, Di, Yi, Cai, Yan, and Lu, Haojie
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4-Hydroxy-2-nonenal (HNE)-modified proteins are closely associated with cellular functions and diseases, so qualitative and quantitative analysis of HNE-modified proteins is very necessary in order to further understand their structures and molecular functions. In this study, we described a six-plex isobaric labeling affinity purification (SiLAP) method based on the interaction of aminoxyTMT six-plex and anti-TMT antibody resin to identify and quantify the HNE modifications simultaneously. The labeling efficiency, ionization efficiency of the aminoxyTMT-tagged peptides, and reliability of the quantification method were investigated in detail. The mass tags were labeled on the modification sites, which could also significantly increase the ionization efficiency, contributing to site-specific identification and quantification of HNE peptides. The SiLAP strategy possessed high sensitivity, accuracy, and good reproducibility to qualitatively and quantitatively analyze HNE-modified proteins/peptides, which could be used to analyze both endogenously and exogenously modified proteins. Using the SiLAP strategy, 2257 HNE-modified peptides mapping 1121 proteins were collectively quantified, which was the largest data set of HNE-modified proteins with detailed modification sites, and 101 proteins were found to be differentially modified by HNE in six liver cell lines. At the same time, 33 endogenously HNE-modified peptides mapping 33 proteins were identified with modification sites.
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- 2019
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14. Normal family of meromorphic functions concerning fixed-points
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Fang, Caiyun and Xu, Yan
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Let $$A>1$$A>1be a constant and $$\mathcal {F}$$Fbe a family of meromorphic functions defined in a domain D. For each $$f\in \mathcal {F}$$f∈F, fhas only zeros of multiplicity at least 3 and satisfies the following conditions: (1) $$|f^{\prime \prime \prime }(z)|\le A|z|$$|f″′(z)|≤A|z|when $$f(z)=0$$f(z)=0; (2) $$f^{\prime \prime \prime }(z)\ne z$$f″′(z)≠z; (3) all poles of fare multiple. In this paper, we characterize the non-normal sequences of $$\mathcal {F}$$F.
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- 2019
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15. Site-Specific Quantification of Protein Palmitoylation by Cysteine-Stable Isotope Metabolic Labeling
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Zhang, Xiaoqin, Zhang, Lei, Ji, Guanghui, Lei, Qunying, Fang, Caiyun, and Lu, Haojie
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Palmitoylation is one of the most important protein translational modifications and plays vital roles in many key biological processes. Aberrant palmitoylation has been associated with a variety of human diseases. So it is of great significance to profile the palmitoylated proteomes qualitatively and quantitatively. Here, we described a novel method based on the cysteine-stable isotope labeling in cell culture (cysteine-SILAC) to facilitate the quantitation of palmitoylated proteins by mass spectrometry (MS), in which “light” or “heavy” samples could be pooled and subjected to the subsequent analysis procedures simultaneously, minimizing systematic errors caused by parallel operations and improving quantitative accuracy and precision. The mass tags lay on the cysteine residues, which were the potential palmitoylated sites, indicating that all the putative modified sites/peptides could be quantified, including the C-terminal peptide of one protein. Due to the isotopically labeled cysteine, the nonspecifically adsorbed peptide without cysteine was singlet in MS spectra, whereas pair peaks should be the signals of putative palmitoylated peptides, which could reduce spectral complexity and achieve double verification for the putative palmitoylated peptides. Finally, the palmitoylome in hepatocellular carcinoma (HCC) cells with different metastasis potentials (MHCC-97L and HCC-LM3 cells) were analyzed for the first time. Totally, 151 proteins were found to be differentially palmitoylated with high confidence, including many important proteins involved in a variety of biological processes, such as protein palmitoylation, cell proliferation, signal transduction, regulation of cell migration, and so on.
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- 2018
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16. Identification of Palmitoylated Transitional Endoplasmic Reticulum ATPase by Proteomic Technique and Pan Antipalmitoylation Antibody
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Fang, Caiyun, Zhang, Xiaoqin, Zhang, Lei, Gao, Xing, Yang, Pengyuan, and Lu, Haojie
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Protein palmitoylation plays a significant role in a wide range of biological processes such as cell signal transduction, metabolism, apoptosis, and carcinogenesis. For high-throughput analysis of protein palmitoylation, approaches based on the acyl-biotin exchange or metabolic labeling of azide/alkynyl-palmitate analogs are commonly used. No palmitoylation antibody has been reported. Here, the palmitoylated proteome of human colon cancer cell lines SW480 was analyzed via a TS-6B-based method. In total, 151 putative palmitoylated sites on 92 proteins, including 100 novel sites, were identified. Except for 3 known palmitoylated transmembrane proteins, ATP1A1, ZDHHC5, and PLP2, some important proteins including kinases, ion channels, receptors, and cytoskeletal proteins were also identified, such as CLIC1, PGK1, PPIA, FKBP4, exportin-2, etc. More importantly, the pan antipalmitoylation antibody was developed and verified for the first time. Our homemade pan antipalmitoylation antiserum could differentiate well protein palmitoylation from mouse brain membrane fraction and SW480 cells, which affords a new technique for analyzing protein palmitoylation by detecting the palmitic acid moiety directly. Furthermore, the candidate protein transitional endoplasmic reticulum ATPase (VCP) identified in SW480 cells was validated to be palmitoylated by Western blotting with anti-VCP antibody and the homemade pan antipalmitoylation antibody.
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- 2016
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17. Isoflurane Protects the Myocardium Against Ischemic Injury via the Preservation of Mitochondrial Respiration and Its Supramolecular Organization
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Lotz, Christopher, Zhang, Jun, Fang, Caiyun, Liem, David, and Ping, Peipei
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Published ahead of print November 7, 2014.
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- 2015
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18. A Novel Average Measure Approach to the Identification of Native-Like Protein Structures Among Decoy Sets
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Li, Juan, Fang, Caiyun, and Fang, Huisheng
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It is a great challenge to predict a protein structure and this challenge has fascinated researchers in different disciplines for many years. Basically the prediction process mainly includes two steps. With the first step that the generation of prediction model increasing fast, the second step that the quality estimation of predicted model i.e. identification of models’ native like structure becomes more and more important. In this study, we developed a simple and effective approach to identify the native-like protein structures among a set of decoys. Three different average measures were used in our study as follows: the average rmsd (armsd), the average alignment score (AAS) and MAXSUB. This approach was evaluated by decoy set (Park-Levitt). Comparison of model quality revealed that a significant correlation existed between these parameters. For example, the average measure could be effectively used to identify native-like protein models. The performance of both armsd and AAS was better than that of clustering. Since many other measures could be used to assess the similarity between protein structures, other analogous approaches might be also useful for the identification of native-like proteins. Finally, data showed that its performance was better than that of other servers in predicting the targets in CASP6, CASP7, CASP9 and CASP10.
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- 2014
19. Multiple technical routes to obtain a proteomics expression profile of French liver samplesElectronic supplementary information (ESI) available. See DOI: 10.1039/c3ay42146e
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Jin, Hong, Zhang, Yang, Xie, Liqi, Shen, Huali, Fang, Caiyun, Lu, Haojie, Gao, Mingxia, Fan, Huizhi, and Yang, Penyuan
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The liver is an important organ and is the biggest digestive gland in the human body. The establishment of a proteome database of the human liver would provide useful information for human liver disease treatment. In order to maximize protein identification and to compare different analyses, multiple technical routes were used for proteome profiling of French liver samples. Five strategies, including two two-dimensional electrophoresis (2DE) methods and three multi-dimensional liquid chromatography (MDLC) methods were evaluated. By using these five routes, 1627 unique proteins were finally identified. Various bioinformatics analyses focused on physicochemical properties and functional classification were used to examine the characteristics of the protein expression profile. Furthermore, comparison of these data with an existing liver expression profile provided by UNIGENE and UNIPROT allowed us to investigate the identification efficiency of this dataset. The different technical methods were evaluated and compared to make a model of the organ proteome profile.
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- 2014
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20. Regulation of Acetylation Restores Proteolytic Function of Diseased Myocardium in Mouse and Human*
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Wang, Ding, Fang, Caiyun, Zong, Nobel C., Liem, David A., Cadeiras, Martin, Scruggs, Sarah B., Yu, Hongxiu, Kim, Allen K., Yang, Pengyuan, Deng, Mario, Lu, Haojie, and Ping, Peipei
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Proteasome complexes play essential roles in maintaining cellular protein homeostasis and serve fundamental roles in cardiac function under normal and pathological conditions. A functional detriment in proteasomal activities has been recognized as a major contributor to the progression of cardiovascular diseases. Therefore, approaches to restore proteolytic function within the setting of the diseased myocardium would be of great clinical significance.
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- 2013
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21. Specific and Reversible Enrichment of Early-Stage Glycated Proteome Based on Thiazolidine Chemistry and Palladium-Mediated Cleavage
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Wu, Linlin, Fei, Weiwei, Liu, Zhiyong, Zhang, Lei, Fang, Caiyun, and Lu, Haojie
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Comprehensive analysis of protein glycation is important for better understanding of its formation mechanism and biological significance. The current preconcentration methods of glycated proteome mainly depend on the reversible combination of boronic acid and cis-dihydroxy group by pH adjustment, but it has inherent limitations (e.g., poor specificity and time-consuming). Herein, for the first time, a novel enrichment method for glycated peptides is proposed based on the reversible chemical reaction between aldehyde and 1,2-aminothiol groups, in which oxidized glycated peptides are captured onto the magnetic nanoparticles via thiazolidine chemistry and then released by palladium-mediated cleavage. The method is rapid, with excellent selectivity (even at a 1:1000 molar ratio of glycated peptides/nonglycated peptides) and high sensitivity (1 fmol/μL). As a good evidence, 1549 glycated peptides were identified from glycated human serum with 94.6% specificity, providing a powerful technique for high-throughput analysis of glycated peptides.
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- 2022
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22. Nuclear proteome profile of C57BL/6J mouse liver
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Zhang, Yang, Fang, CaiYun, Bao, HuiMin, Fan, HuiZhi, Shen, HuaLi, and Yang, PengYuan
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The liver proteome can serve as a reference to better understand both disease mechanisms and possible therapeutics, since the liver is an important organ in the body that performs a large number of tasks. Here we identify the organelle proteome of C57BL/6J mouse liver nuclei as a promising strategy to enrich low abundance proteins, in the sense that analysis of whole liver cells is rather complex for current techniques and may not be suitable for proteins with low abundance. Evaluation of nucleus integrity and purity was performed to demonstrate the effectiveness of the optimized isolation procedure. The extracted nuclear proteins were identified by 2-DE MS analyses, and a total of 748 proteins were identified. Bioinformatic analyses were performed to demonstrate the physicochemical properties, cellular locations and functions of the proteins.
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- 2013
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23. Proteome Dynamics and Proteome Function of Cardiac 19S Proteasomes
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Wang, Ding, Zong, Chenggong, Koag, Myong-chul, Wang, Yueju, Drews, Oliver, Fang, Caiyun, Scruggs, Sarah B., and Ping, Peipei
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Myocardial proteasomes are comprised of 20S core particles and 19S regulatory particles, which together carry out targeted degradation of cardiac proteins. The 19S complex is unique among the regulators of proteasomes in that it affects both the capacity and specificity of protein degradation. However, a comprehensive molecular characterization of cardiac 19S complexes is lacking. In this investigation, we tailored a multidimensional chromatography-based purification strategy to isolate structurally intact and functionally viable 19S complexes from murine hearts. Two distinct subpopulations of 19S complexes were isolated based upon (1) potency of activating 20S proteolytic activity, and (2) molecular composition using a combination of immuno-detection, two-dimensional-differential gel electrophoresis, and MS-based approaches. Heat shock protein 90 (Hsp90) was identified to be characteristic to 19S subpopulation I. The physical interaction of Hsp90 with 19S complexes was demonstrated via multiple approaches. Inhibition of Hsp90 activity using geldanamycin or BIIB021 potentiated the ability of subpopulation I to activate 20S proteasomes in the murine heart, thus demonstrating functional specificity of Hsp90 in subpopulation I. This investigation has advanced our understanding of the molecular heterogeneity of cardiac proteasomes by identifying molecularly and functionally distinct cardiac 19S complexes. The preferential association of Hsp90 with 19S subpopulation I unveils novel targets for designing proteasome-based therapeutic interventions for combating cardiac disease.
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- 2011
24. Proteome Dynamics and Proteome Function of Cardiac 19S Proteasomes*
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Wang, Ding, Zong, Chenggong, Koag, Myong-chul, Wang, Yueju, Drews, Oliver, Fang, Caiyun, Scruggs, Sarah B., and Ping, Peipei
- Abstract
Myocardial proteasomes are comprised of 20S core particles and 19S regulatory particles, which together carry out targeted degradation of cardiac proteins. The 19S complex is unique among the regulators of proteasomes in that it affects both the capacity and specificity of protein degradation. However, a comprehensive molecular characterization of cardiac 19S complexes is lacking. In this investigation, we tailored a multidimensional chromatography-based purification strategy to isolate structurally intact and functionally viable 19S complexes from murine hearts. Two distinct subpopulations of 19S complexes were isolated based upon (1) potency of activating 20S proteolytic activity, and (2) molecular composition using a combination of immuno-detection, two-dimensional-differential gel electrophoresis, and MS-based approaches. Heat shock protein 90 (Hsp90) was identified to be characteristic to 19S subpopulation I. The physical interaction of Hsp90 with 19S complexes was demonstrated via multiple approaches. Inhibition of Hsp90 activity using geldanamycin or BIIB021 potentiated the ability of subpopulation I to activate 20S proteasomes in the murine heart, thus demonstrating functional specificity of Hsp90 in subpopulation I. This investigation has advanced our understanding of the molecular heterogeneity of cardiac proteasomes by identifying molecularly and functionally distinct cardiac 19S complexes. The preferential association of Hsp90 with 19S subpopulation I unveils novel targets for designing proteasome-based therapeutic interventions for combating cardiac disease.
- Published
- 2011
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