A novel approach for identifying the heme-binding proteins from mouse tissues.
- Author:
Xiaolei LI
1
;
Xiaoshan WANG
;
Kang ZHAO
;
Zhengfeng ZHOU
;
Caifeng ZHAO
;
Ren YAN
;
Liang LIN
;
Tingting LEI
;
Jianning YIN
;
Rong WANG
;
Zhongsheng SUN
;
Zuyuan XU
;
Jingyue BAO
;
Xiuqing ZHANG
;
Xiaoli FENG
;
Siqi LIU
Author Information
1. Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Carrier Proteins;
biosynthesis;
genetics;
Electrophoresis, Gel, Two-Dimensional;
Electrophoresis, Polyacrylamide Gel;
Heme;
chemistry;
Hemeproteins;
biosynthesis;
genetics;
Mass Spectrometry;
Mice;
Mice, Inbred ICR;
Protein Binding;
Proteins;
chemistry;
Proteome;
Proteomics;
methods;
Sepharose;
chemistry;
Tissue Distribution
- From:
Genomics, Proteomics & Bioinformatics
2003;1(1):78-86
- CountryChina
- Language:English
-
Abstract:
Heme is a key cofactor in aerobic life, both in eukaryotes and prokaryotes. Because of the high reactivity of ferrous protoporphyrin IX, the reactions of heme in cells are often carried out through heme-protein complexes. Traditionally studies of heme-binding proteins have been approached on a case by case basis, thus there is a limited global view of the distribution of heme-binding proteins in different cells or tissues. The procedure described here is aimed at profiling heme-binding proteins in mouse tissues sequentially by 1) purification of heme-binding proteins by heme-agarose, an affinity chromatographic resin; 2) isolation of heme-binding proteins by SDS-PAGE or two-dimensional electrophoresis; 3) identification of heme-binding proteins by mass spectrometry. In five mouse tissues, over 600 protein spots were visualized on 2-DE gel stained by Commassie blue and 154 proteins were identified by MALDI-TOF, in which most proteins belong to heme related. This methodology makes it possible to globally characterize the heme-binding proteins in a biological system.
