A Novel Approach for Identifying the Heme—Binding Proteins from Mouse Tissues

• Published in: Genomics, proteomics & bioinformatics proteomics & bioinformatics, 2003-02, Vol.1 (1), p.78-86
• Main Authors: Li, Xiaolei, Wang, Xiaoshan, Zhao, Kang, Zhou, Zhengfeng, Zhao, Caifeng, Yan, Ren, Lin, Liang, Lei, Tingting, Yin, Jianning, Wang, Rong, Feng, Xiaoli, Liu, Siqi
• Format: Article
• Language: English
• Subjects: Animals; Carrier Proteins - biosynthesis; Carrier Proteins - genetics; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; heme; Heme - chemistry; heme-agarose; Hemeproteins - biosynthesis; Hemeproteins - genetics; Mass Spectrometry; Methods; Mice; Mice, Inbred ICR; mouse; Protein Binding; Proteins - chemistry; Proteome; proteomics; Proteomics - methods; Sepharose - chemistry; Tissue Distribution; 亚铁血红素; 亚铁血红素－琼脂糖; 亚铁血红素－蛋白质复合体; 鉴别方法
• ISSN: 1672-0229; 2210-3244
• DOI: 10.1016/S1672-0229(03)01011-8

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 hemebinding 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 hemeagarose,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 2DE 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.