title: Native protein mapping and visualization of protein interactions in the area of human plasma high-density lipoprotein by combining nondenaturing micro 2DE and quantitative LC-MS/MS.
abstract: A human plasma sample was subjected to nondenaturing micro 2DE and a gel area (5 mm × 18 mm) that includes high-density lipoprotein (HDL) was cut into 1 mm × 1 mm squares, then the proteins in the 90 gel pieces were analyzed by quantitative LC-MS/MS. Grid-cutting of the gel was employed to; (i) ensure the total analysis of the proteins in the area, (ii) standardize the conditions of analysis by LC-MS/MS, (iii) reconstruct the protein distribution patterns from the quantity data. Totally 154 proteins were assigned in the 90 gel pieces 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. These results for the first time visualized the localization of HDL apolipoproteins on a nondenaturing 2DE gel and strongly suggested their interactions.
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, P. R. China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Jin
authors.forename: Shujie
authors.initials: S
authors.lastname: Bu
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Yuan
authors.forename: Takashi
authors.initials: T
authors.lastname: Manabe
authors.forename: Wen
authors.initials: W
authors.lastname: Tan
mesh_terms: D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D008075:Lipoproteins, HDL / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001798:Blood Proteins; D008075:Lipoproteins, HDL
keywords: High-density lipoprotein; Human plasma; Nondenaturing micro 2DE; Protein-protein interaction; Quantitative LC-MS/MS