Zhang Qibin, Tang Ning, Schepmoes Athena A, Phillips Lawrence S, Smith Richard D, Metz Thomas O
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
J Proteome Res. 2008 May;7(5):2025-32. doi: 10.1021/pr700763r. Epub 2008 Apr 9.
Nonenzymatic glycation of peptides and proteins by d-glucose has important implications in the pathogenesis of diabetes mellitus, particularly in the development of diabetic complications. In this work, we report the first proteomics-based characterization of nonenzymatically glycated proteins in human plasma and erythrocyte membranes from individuals with normal glucose tolerance, impaired glucose tolerance, and type 2 diabetes mellitus. Phenylboronate affinity chromatography was used to enrich glycated proteins and glycated tryptic peptides from both human plasma and erythrocyte membranes. The enriched peptides were subsequently analyzed by liquid chromatography coupled with electron transfer dissociation-tandem mass spectrometry, resulting in the confident identification of 76 and 31 proteins from human plasma and erythrocyte membranes, respectively. Although most of the glycated proteins could be identified in samples from individuals with normal glucose tolerance, slightly higher numbers of glycated proteins and more glycation sites were identified in samples from individuals with impaired glucose tolerance and type 2 diabetes mellitus.
D-葡萄糖对肽和蛋白质的非酶糖基化作用在糖尿病发病机制中具有重要意义，尤其是在糖尿病并发症的发展过程中。在本研究中，我们首次基于蛋白质组学对葡萄糖耐量正常、葡萄糖耐量受损和2型糖尿病患者的人血浆及红细胞膜中的非酶糖基化蛋白质进行了表征。采用苯基硼酸亲和色谱法从人血浆和红细胞膜中富集糖基化蛋白质和糖基化胰蛋白酶肽段。随后，通过液相色谱与电子转移解离串联质谱联用对富集的肽段进行分析，分别从人血浆和红细胞膜中可靠鉴定出76种和31种蛋白质。虽然在葡萄糖耐量正常个体的样本中可鉴定出大多数糖基化蛋白质，但在葡萄糖耐量受损和2型糖尿病患者的样本中鉴定出的糖基化蛋白质数量略多，糖基化位点也更多。
