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Aims: Adolescent offspring exposed to maternal diabetes during intrauterine life show a less favourable metabolic profile than the background population. Here, we hypothesize that offspring of women with type 1 diabetes (T1D), possess sex‐specific alterations in the serum profile of proteins involved in lipid, metabolic and transport processes and that these alterations are associated with lipid profile and indices of insulin sensitivity and secretion. Methods: A prospective nationwide follow‐up study (EPICOM) in a Danish population. Blood samples were assessed from offspring of women with T1D (index offspring, n = 267, 13–20 years), and matched control offspring (n = 290). Serum proteins were analysed using a 25‐plex cardio‐metabolic targeted proteomics assay, which includes 12 apolipoproteins and 13 transport and inflammatory proteins. Results: Apolipoprotein D (ApoD) and transthyretin (TTR) were reduced in index females as compared to female controls (−8.1%, p < 0.001 and −6.1%, p = 0.006 respectively), but not in index males (2.2%, p = 0.476 and −2.4%, p = 0.731 respectively). Sex‐dependent inverse associations between exposure to maternal T1D in utero and ApoD and TTR were significant after adjusting for age, BMI‐SDS and Tanner stage (OR = 0.252 [95% CI 0.085, 0.745], p = 0.013 and OR = 0.149 [95% CI 0.040, 0.553], p = 0.004). ApoD correlated to indices of insulin sensitivity and secretion in a similar sex‐specific pattern in crude and adjusted analyses. Conclusions: Low ApoD may be regarded as an early risk marker of metabolic syndrome. A possible link between ApoD and cardiovascular disease needs further investigation. [ABSTRACT FROM AUTHOR]

Background: Gestational diabetes mellitus (GDM) is a common pregnancy complication associated with adverse outcomes including preeclampsia, caesarean section, macrosomia, neonatal morbidity and future development of type 2 diabetes in both mother and child. Current selective screening strategies rely on clinical risk factors such as age, family history of diabetes, macrosomia or GDM in a previous pregnancy, and they possess a relatively low specificity. Here we hypothesize that novel first trimester protein predictors of GDM can contribute to the current selective screening strategies for early and accurate prediction of GDM, thus allowing for timely interventions. Methods: A proteomics discovery approach was applied to first trimester sera from obese (BMI ≥27 kg/m2) women (n = 60) in a nested case-control study design, utilizing tandem mass tag labelling and tandem mass spectrometry. A subset of the identified protein markers was further validated in a second set of serum samples (n = 210) and evaluated for their contribution as predictors of GDM in relation to the maternal risk factors, by use of logistic regression and receiver operating characteristic analysis. Results: Serum proteomic profiling identified 25 proteins with significantly different levels between cases and controls. Three proteins; afamin, serum amyloid P-component and vitronectin could be further confirmed as predictors of GDM in a validation set. Vitronectin was shown to contribute significantly to the predictive power of the maternal risk factors, indicating it as a novel independent predictor of GDM. Conclusions: Current selective screening strategies can potentially be improved by addition of protein predictors. [ABSTRACT FROM AUTHOR]

Food allergies mediated by specific IgE (sIgE) have a significant socioeconomic impact on society. Evaluating the IgE cross-reactivity between allergens from different allergen sources can enable the better management of these potentially life-threatening adverse reactions to food proteins and enhance food safety. A novel banana fruit allergen, S-adenosyl-L-homocysteine hydrolase (SAHH), has been recently identified and its recombinant homolog was heterologously overproduced in E. coli. In this study, we performed a search in the NCBI (National Center for Biotechnology Information) for SAHH homologs in ryegrass, latex, and kiwifruit, all of which are commonly associated with pollen-latex-fruit syndrome. In addition, Western immunoblot analysis was utilized to identify the cross-reactive IgE to banana SAHH in the sera of patients with a latex allergy, kiwifruit allergy, and ryegrass allergy. ClustalOmega analysis showed more than 92% amino acid sequence identity among the banana SAHH homologs in ryegrass, latex, and kiwifruit. In addition to five B-cell epitopes, in silico analysis predicted eleven T-cell epitopes in banana SAHH, seventeen in kiwifruit SAHH, twelve in ryegrass SAHH, and eight in latex SAHH, which were related to the seven-allele HLA reference set (HLA-DRB1*03:01, HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DRB4*01:01, HLA-DRB5*01:01). Four T-cell epitopes were identical in banana and kiwifruit SAHH (positions 328, 278, 142, 341), as well as banana and ryegrass SAHH (positions 278, 142, 96, and 341). All four SAHHs shared two T-cell epitopes (positions 278 and 341). In line with the high amino acid sequence identity and B-cell epitope homology among the analyzed proteins, the cross-reactive IgE to banana SAHH was detected in three of three latex-allergic patients, five of six ryegrass-allergic patients, and two of three kiwifruit-allergic patients. Although banana SAHH has only been studied in a small group of allergic individuals, it is a novel cross-reactive food allergen that should be considered when testing for pollen-latex-fruit syndrome. [ABSTRACT FROM AUTHOR]

Sulfur-containing amino acid residues function in antioxidative responses, which can be induced by the reactive oxygen species generated by excessive copper and hydrogen peroxide. In all Na+/K+, Ca2+, and H+ pumping P-type ATPases, a cysteine residue is present two residues upstream of the essential aspartate residue, which is obligatorily phosphorylated in each catalytic cycle. Despite its conservation, the function of this cysteine residue was hitherto unknown. In this study, we analyzed the function of the corresponding cysteine residue (Cys-327) in the autoinhibited plasma membrane H+-ATPase isoform 2 (AHA2) from Arabidopsis thaliana by mutagenesis and heterologous expression in a yeast host. Enzyme kinetics of alanine, serine, and leucine substitutions were identical with those of the wild-type pump but the sensitivity of the mutant pumps was increased towards copper and hydrogen peroxide. Peptide identification and sequencing by mass spectrometry demonstrated that Cys-327 was prone to oxidation. These data suggest that Cys-327 functions as a protective residue in the plasma membrane H+- ATPase, and possibly in other P-type ATPases as well. [ABSTRACT FROM AUTHOR]

The mechanisms of one-electron protein oxidation are complicated and still not well-understood. In this work, we investigated the reaction of sensitized photo-oxidation using carboxybenzophenone (CB) as a sensitizer and a PR-10 protein (MtN13) as a quencher, which is intrinsically complicated due to the complex structure of the protein and multiple possibilities of CB attack. To predict and examine the possible reactions precisely, the 3D structure of the MtN13 protein was taken into account. Our crystallographic studies revealed a specific binding of the CB molecule in the protein's hydrophobic cavity, while mass spectrometry identified the amino acid residues (Met, Tyr, Asp and Phe) creating adducts with the sensitizer, thus indicating the sites of 3CB* quenching. In addition, protein aggregation was also observed. The detailed mechanisms of CB quenching by the MtN13 molecule were elucidated by an analysis of transient products by means of time-resolved spectroscopy. The investigation of the transient and stable products formed during the protein photo-oxidation was based on the data obtained from HPLC-MS analysis of model compounds, single amino acids and dipeptides. Our proposed mechanisms of sensitized protein photo-oxidation emphasize the role of a ground state complex between the protein and the sensitizer and indicate several new and specific products arising as a result of one-electron oxidation. Based on the analysis of the transient and stable products, we have demonstrated the influence of neighboring groups, especially in the case of Tyr oxidation, where the tyrosyl radical can be formed via a direct electron transfer from Tyr to CB* or via an intramolecular electron transfer from Tyr to Met radical cation Met > S●+ or thiyl radical CysS● from neighboring oxidized groups. [Display omitted] • Aromatic sensitizers can form the ground state complex with proteins via interactions with Phe residues. • The complex protein-sensitizer influences the protein oxidation processes. • Electron transfer from Tyr to radicals on Met in protein models occurs as in model compounds yielding TyrO●. • The stable products of oxidation were identified on amino acid side chain that are not prone to oxidation, e.g. Asp. • The mechanism of photo-oxidation of MtN13 was described by time resolved methods and UPLC-MS.. [ABSTRACT FROM AUTHOR]