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Investigation of Non-Enzymatic Glycosylation of Human Serum Albumin Using Ion Trap-Time of Flight Mass Spectrometry
National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China
State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong 250100, China
Life Science and Clinical Department, Shimadzu International Incorporation, Beijing 100020, China
Division of Endocrinology and Metabolism, Provincial Hospital affiliated to Shandong University, Jinan, Shandong 250021, China
* Authors to whom correspondence should be addressed.
Received: 4 June 2012; in revised form: 8 July 2012 / Accepted: 13 July 2012 / Published: 25 July 2012
Abstract: 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.
Keywords: glycation; human serum albumin; ion trap-time of flight; mass spectrometry; peptide fingerprinting; diabetes; incorporation ratio
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Cite This Article
MDPI and ACS Style
Bai, X.; Wang, Z.; Huang, C.; Wang, Z.; Chi, L. Investigation of Non-Enzymatic Glycosylation of Human Serum Albumin Using Ion Trap-Time of Flight Mass Spectrometry. Molecules 2012, 17, 8782-8794.
Bai X, Wang Z, Huang C, Wang Z, Chi L. Investigation of Non-Enzymatic Glycosylation of Human Serum Albumin Using Ion Trap-Time of Flight Mass Spectrometry. Molecules. 2012; 17(8):8782-8794.
Bai, Xue; Wang, Zhangjie; Huang, Chengcai; Wang, Zhe; Chi, Lianli. 2012. "Investigation of Non-Enzymatic Glycosylation of Human Serum Albumin Using Ion Trap-Time of Flight Mass Spectrometry." Molecules 17, no. 8: 8782-8794.