Wang H.,Tongji University |
Feng L.,Tongji University |
Hu J.,Shanghai Applied Protein Technology Co. |
Xie C.,Tongji University |
Wang F.,Tongji University
Experimental Eye Research | Year: 2013
Proliferative diabetic retinopathy (PDR) is a serious microangiopathic complication of diabetes mellitus and a major cause of blindness in working-age adults. Diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. Therefore, we performed a comprehensive proteomic analysis and compared the protein profiles of vitreous humor from type 2 diabetic patients with PDR (n = 8) and that from normal human eyes donated for corneal transplant (n = 8). Using reversed phase high-performance liquid chromatography (RP-HPLC) coupled to electrospray Ionization tandem mass spectrometry (ESI-MS/MS), we identified 96 significant differentially expressed proteins (abundance ratio > 1.5, p < 0.05), including 37 and 59 proteins up- and downregulated in PDR vitreous compared with the control, respectively. Biological pathway analysis revealed 44 proteins involved in 56 biological pathways; among them, the most remarkable pathways differentially represented between PDR and normal vitreous were the glycolysis/gluconeogenesis, complement and coagulation cascades, gap junction, and phagosome pathways. The differential expressions of angiopoietin-related protein 6, apolipoprotein A-I, estrogen receptor alpha, and tubulin were confirmed by western blot analysis. These data provide insight into the molecular events possibly involved in the pathogenesis of PDR and widen the scope of potential avenues for new therapies for PDR. © 2012 Elsevier Ltd.
Wu L.,Henan Agricultural University |
Wu L.,Key Laboratory of Physiological Ecology and Genetic Improvement of Food Crops in Henan Province |
Wang S.,Henan Agricultural University |
Wang S.,Key Laboratory of Physiological Ecology and Genetic Improvement of Food Crops in Henan Province |
And 9 more authors.
Amino Acids | Year: 2015
Protein phosphorylation plays a pivotal role in the regulation of many cellular events. No information is yet available, however, on protein phosphorylation in plants in response to virus infection. In this study, we characterized phosphoproteomes of resistant and susceptible genotypes of maize (Zea mays L.) in response to Sugarcane mosaic virus (SCMV) infection. Based on isotope tags for relative and absolute quantification technology, TiO2 enrichment method and LC-MS/MS analysis, we identified 65 and 59 phosphoproteins respectively, whose phosphorylation level regulated significantly in susceptible and resistant plants. Some identified phosphoproteins were shared by both genotypes, suggesting a partial overlapping of the responsive pathways to virus infection. While several phosphoproteins are well-known pathogen response phosphoproteins, virus infection differentially regulates most other phosphoproteins, which has not been reported in literature. Changes in protein phosphorylation status indicated that response to SCMV infection encompass a reformatting of major cellular processes. Our data provide new valuable insights into plant-virus interactions. © 2014 Springer-Verlag Wien.
Yao Z.,Shenyang University |
Yao Z.,Liaoning Medical University |
Yu H.,ZhongXin Biotechnology Shanghai Co. |
Xuan D.,Uniformed Services University of the Health Sciences |
And 3 more authors.
Current Eye Research | Year: 2010
Purpose: To investigate proteomic profiles of normal human lenses and their key proteins in proteinprotein interactions (PPIs). Materials and Methods: Water-soluble and water-insoluble proteins extracted from human lenses were first separated by one-dimensional sodium dodecyl sulfate polyacrylamide gel, and then in-gel digested with trypsin into peptides eluted by reversed-phase high-performance liquid chromatography. The eluted peptides were analyzed by linear ion trap tandem mass spectrometry (MS/MS). The raw data was filtered by TurboSEQUEST algorithm. The reverse database was used for peptide false-positive rate estimation. A network chart was constructed by the identified lens PPIs in accordance with interaction database systems. Results: From normal human lenses 339 proteins in total were identified, including many formerly unidentified low-abundance proteins. Key proteins we recognized included plectin, actin, spectrin (α, β), vimentin, 14-3-3 protein (β/α, ζ/δ, ε, γ, η), TSC2, guanine nucleotide-releasing protein, laminin γ, mitogen-activated protein kinase, α-A- crystallin, heat-shock protein (α, β), glyceraldehyde 3-phosphate dehydrogenase, and collagen IV α. Conclusions: Key proteins of normal human lenses were studied by constructing a network chart of the identified lens PPIs. The results suggest that linear ion trap MS/MS is an effective tool for detecting low-abundance proteins of human lenses. This study provides valuable data for further proteomic research of the human lens development and lens diseases. © 2010 Informa Healthcare USA, Inc.
Wang H.,Tongji University |
Feng L.,Tongji University |
Hu J.W.,Shanghai Applied Protein Technology Co. |
Xie C.L.,Tongji University |
Wang F.,Tongji University
Proteome Science | Year: 2012
Background: Diabetes can lead to serious microvascular complications such as proliferative diabetic retinopathy (PDR), which results in severe vision loss. The diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. The vitreous humour can be utilised in a variety of studies aimed toward the discovery of new targets for the treatment or prevention of PDR and the identification of novel disease mechanisms. The aim of this study was to compare the protein profile of vitreous humour from diabetic patients with PDR with that of vitreous humour from normal human eyes donated for corneal transplant.Results: Vitreous humour from type 2 diabetic patients with PDR (n = 10) and from normal human eyes donated for corneal transplant (n = 10) were studied. The comparative proteomic analysis was performed using two-dimensional fluorescence difference gel electrophoresis (2-D DIGE). Differentially produced proteins (abundance ratio > 2 or < -2, p < 0.01) were identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF tandem mass spectrometry. A total of 1242 protein spots were detected on the 2-D master gel of the samples, and 57 spots that exhibited statistically significant variations were successfully identified. The spots corresponded to peptide fragments of 29 proteins, including 8 proteins that increased and 21 proteins that decreased in PDR. Excluding the serum proteins from minor vitreous haemorrhage, 19 proteins were found to be differentially produced in PDR patients compared with normal subjects; 6 of these proteins have never been reported to be differentially expressed in PDR vitreous: N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 (DDAH 1), tubulin alpha-1B chain, gamma-enolase, cytosolic acyl coenzyme A thioester hydrolase, malate dehydrogenase and phosphatidylethanolamine-binding protein 1 (PEBP 1). The differential production of pigment epithelium-derived factor (PEDF) and clusterin was confirmed by Western blot analysis.Conclusions: These data provide an in-depth analysis of the human vitreous proteome and reveal protein alterations that are possibly involved in the pathogenesis of PDR. Further investigation of these special proteins may provide potential new targets for the treatment or the prevention of PDR. © 2012 Wang et al; licensee BioMed Central Ltd.
Li L.,Fudan University |
Li L.,Shanghai Applied Protein Technology Co. |
Jiao J.,Fudan University |
Cai Y.,Fudan University |
And 2 more authors.
Analytical Chemistry | Year: 2015
The sensitive and specific detection of glycans via mass spectrometry (MS) remains a significant challenge due to their low abundance in complex biological mixtures, inherent lack of hydrophobicity, and suppression by other, more abundant biological molecules (proteins/peptides) or contaminants. A new strategy for the sensitive and selective MS analysis of glycans based on fluorous chemistry is reported. Glycan reducing ends were derivatized with a hydrophobic fluorinated carbon tag, increasing glycan ionization efficiency during MS by more than an order of magnitude. More importantly, the fluorinated carbon tag enabled efficient fluorous solid-phase extraction (FSPE) to specifically enrich the glycans from contaminated solutions and protein mixtures. Finally, we successfully analyzed the N-glycome in human serum using this new method. © 2015 American Chemical Society.