Dataset Information

Proteomic analysis of cellular soluble proteins from human bronchial smooth muscle cells by combining nondenaturing micro 2DE and quantitative LC-MS/MS: Preparation of more than 4000 native protein maps

ABSTRACT: Soluble proteins of human bronchial smooth muscle cells (HBSMC) were separated by nondenaturing micro 2DE and a 30 mm X 40 mm area of the CBB-stained slab gel (1.0 mm thick) was cut into 1.1 mm X 1.1 mm squares, then the proteins in the 972 gel pieces (squares) were applied to 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 [Jin, et al. Electrophoresis 2014, 35, 2055-2064, PRIDE Project PXD000852]. Totally 4323 proteins were identified in successfully analyzed 967 squares and the quantity distribution of each was reconstructed as a color density pattern (a native protein map). The quantity of the proteins distributed from 3.6 % to 1 X 10-5 % of the total protein quantity in the grid area. Each protein map was characterized by many features, such as the position of quantity peak square, number of detected squares, degree of concentration (focused or dispersed), etc. About 4 percent of the proteins were detected in 100 or more squares, suggesting that they might be ubiquitous and interacting with other proteins. In contrast, many proteins showed more concentrated quantity distribution and the quantity peak positions of 565 proteins with defined degree of concentration were summarized into a quantity peak map. These results for the first time visualized the distribution patterns of cellular proteins on a nondenaturing 2DE gel, which would further provide information on their interactions.

INSTRUMENT(S): Synapt G2-S HDMS

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr Ya Jin

PROVIDER: MSV000080002 | MassIVE | Sat Jul 23 05:01:00 BST 2016

SECONDARY ACCESSION(S): PXD001471

REPOSITORIES: MassIVE

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Proteomic analysis of cellular soluble proteins from human bronchial smooth muscle cells by combining nondenaturing micro 2DE and quantitative LC-MS/MS. 1. Preparation of more than 4000 native protein maps.

Jin Ya Y Zhang Jun J Yuan Qi Q Manabe Takashi T Tan Wen W

Electrophoresis 20150617 15

Soluble proteins of human bronchial smooth muscle cells (HBSMC) were separated by nondenaturing micro 2DE and a 30 mm × 40 mm area of the CBB-stained slab gel (1.0 mm thick) was cut into 1.1 mm × 1.1 mm squares, then the proteins in the 972 gel pieces (squares) were applied to 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 ...[more]

PMID: 25931155

Publication: 1/2

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