Investigation of Non-Enzymatic Glycosylation of Human Serum Albumin Using Ion Trap-Time of Flight Mass Spectrometry

被引:30
|
作者
Bai, Xue [2 ]
Wang, Zhangjie [1 ]
Huang, Chengcai [3 ]
Wang, Zhe [4 ]
Chi, Lianli [1 ,2 ]
机构
[1] Shandong Univ, State Key Lab Microbial Technol, Jinan 250100, Shandong, Peoples R China
[2] Shandong Univ, Natl Glycoengn Res Ctr, Jinan 250100, Shandong, Peoples R China
[3] Shimadzu Int Inc, Life Sci & Clin Dept, Beijing 100020, Peoples R China
[4] Shandong Univ, Prov Hosp, Div Endocrinol & Metab, Jinan 250100, Shandong, Peoples R China
关键词
glycation; human serum albumin; ion trap-time of flight; mass spectrometry; peptide fingerprinting; diabetes; incorporation ratio; PERFORMANCE LIQUID-CHROMATOGRAPHY; GLYCATED ALBUMIN; AFFINITY-CHROMATOGRAPHY; IDENTIFICATION; PROTEIN; SYSTEM; ELECTROPHORESIS; SITE;
D O I
10.3390/molecules17088782
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Non-enzymatic glycosylation or glycation involves covalent attachment of reducing sugar residues to proteins without enzyme participation. Glycation of glucose to human serum albumin in vivo is related to diabetes and many other diseases. We present an approach using liquid chromatography coupled to an electrospray ionization source of a hybrid ion trap-time of flight (IT-TOF-MS/MS) tandem mass spectrometer to identify the glycation sites on serum albumin from both a healthy person and a diabetic patient. The MetID software, which is commonly used for screening metabolites, is adapted for peptide fingerprinting based on both m/z values and isotopic distribution profiles. A total of 21 glycation sites from the healthy person and 16 glycation sites from the diabetic patient were identified successfully. We also demonstrate the use of matrix assisted laser desorption ionization-time of flight mass spectrometry to estimate the incorporation ratio of glucose to albumin during glycation. Results from this study show that the glycation in healthy person is more complicated than previously thought. Further analysis of incorporation ratio distribution may be necessary to accurately reflect the change of serum albumin glycation in diabetic patients.
引用
收藏
页码:8782 / 8794
页数:13
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