title: A novel approach for identifying the heme-binding proteins from mouse tissues.
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.
authors.affiliation: Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Li
authors.forename: Xiaoshan
authors.initials: X
authors.lastname: Wang
authors.forename: Kang
authors.initials: K
authors.lastname: Zhao
authors.forename: Zhengfeng
authors.initials: Z
authors.lastname: Zhou
authors.forename: Caifeng
authors.initials: C
authors.lastname: Zhao
authors.forename: Ren
authors.initials: R
authors.lastname: Yan
authors.forename: Liang
authors.initials: L
authors.lastname: Lin
authors.forename: Tingting
authors.initials: T
authors.lastname: Lei
authors.forename: Jianning
authors.initials: J
authors.lastname: Yin
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
authors.forename: Zhongsheng
authors.initials: Z
authors.lastname: Sun
authors.forename: Zuyuan
authors.initials: Z
authors.lastname: Xu
authors.forename: Jingyue
authors.initials: J
authors.lastname: Bao
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Feng
authors.forename: Siqi
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000737:chemistry; D000081062:Heme-Binding Proteins; D006420:Hemeproteins / Q000096:biosynthesis* / Q000235:genetics*; D013058:Mass Spectrometry; D051379:Mice; D008813:Mice, Inbred ICR; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods; D012685:Sepharose / Q000737:chemistry; D014018:Tissue Distribution
chemical_list: D002352:Carrier Proteins; D000081062:Heme-Binding Proteins; D006420:Hemeproteins; D011506:Proteins; D020543:Proteome; D006418:Heme; D012685:Sepharose