Proteomic profiling of nonenzymatically glycated proteins in human plasma and erythrocyte membranes.
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.
Zhang
Qibin
Q
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Tang
Ning
N
Schepmoes
Athena A
AA
Phillips
Lawrence S
LS
Smith
Richard D
RD
Metz
Thomas O
TO
D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000032:analysis* / Q000235:genetics; D003924:Diabetes Mellitus, Type 2 / Q000097:blood; D004910:Erythrocyte Membrane / Q000737:chemistry*; D020869:Gene Expression Profiling; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000235:genetics; D010949:Plasma / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D011897:Random Allocation
D001798:Blood Proteins; D010455:Peptides; D020543:Proteome