Profiling of human HDL and LDL associated proteins by native 2DE-MS mapping
作者单位：Institute of Biomedical and Pharmaceutical SciencesGuangdong University of Technology School of Bioscience and BioengineeringSouth ChinaUniversity of Technology Faculty of ScienceEhime University
会议名称：《2018年中国质谱学术大会（CMSC2018）》
会议日期：2018年
学科分类：081704[工学-应用化学] 07[理学] 1001[医学-基础医学(可授医学、理学学位)] 08[工学] 0817[工学-化学工程与技术] 070302[理学-分析化学] 0703[理学-化学] 10[医学]
关 键 词：human lipoproteins HDL LDL nondenaturing 2DE LC-MS/MS mapping
摘 要：Plasma lipoproteins are soluble complexes of proteins and lipids that transport lipids in the circulation. Historically, lipoproteins were categorized and named by density range, thus each class contains a group of molecules varying in size, charges and lipid and protein compositions. Such differences were demonstrated to contribute to their specific physiological functions and pathological roles. However, profiling of the protein content and interactions of lipoproteins has always been analytically challenging and requires methods of high sensitivity and high resolution. In our work, a method employing nondenaturing 2 DE, grid gel-cutting and quantitative LC-MS/MS has been used to analyze the associated proteins in human plasma high-density lipoprotein(HDL) and low-density lipoprotein(LDL) [1,2]. A human plasma sample was separated by nondenaturing 2 DE, the gel areas that were detected by immunochemical staining to contain HDL and LDL were respectively subjected for grid cut. For HDL, the 5 mm × 18 mm area was cut into 90 squares of 1 mm × 1 mm, and each of the gel piece was analyzed in standardized procedures of in-gel digestion and quantitative LC-MS/MS. Totally 154 proteins were detected and the quantity distribution of each was reconstructed as a color-density pattern(a native protein map). The map of apolipoprotein(Apo) A-I showed a wide apparent mass distribution characteristic to HDL and was compared with the maps of the other 153 proteins. Eleven proteins showed maps of wide distribution that overlapped with the map of Apo A-I, and all have been reported to be the components of HDL. Further, seven minor proteins associated with HDL were detected at the gel positions of high Apo A-I quantity. In the analysis of LDL, the 18 mm × 4.8 mm area was cut into 72 squares. A total of 253 proteins were detected and their native maps were reconstructed. Apo B-100 was shown to be the most abundant protein in the grid-cut area, concentrated at p I ca. 5.4–6.1 and appare