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- Title
The Functional Role of Arginine 901 at the C-Terminus of the Human Anion Transporter Band 3 Protein.
- Authors
Takazaki, Shinya; Abe, Yoshito; Donchon Kang; Chunyan Li; Xiuri Jin; Ueda, Tadashi; Hamasaki, Naotaka
- Abstract
To determine which arginine residues are responsible for band 3-mediated anion transport, we analyzed hydroxyphenylglyoxal (HPG)-modified band 3 protein in native erythrocyte membranes. HPG-modification leads to inhibition of the transport of phosphoenolpyruvate, a substrate for band 3–mediated transport. We analyzed the HPG-modified membranes by reverse phase-HPLC, and determined that arginine 901 was modified by HPG. To determine the role of Arg 901 in the conformational change induced by anion exchange, we analyzed HPG-modification of the membranes when 4,4′-dinitrostilbene-2,2′-disulfonic acid (DNDS) or diethypyrocarbonate (DEPC) was present. DNDS and DEPC fix band 3 in the outward and inward conformations, respectively. HPG-modification was unaffected in the presence of DEPC but decreased in the presence of DNDS. In addition to that, 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), which specifically reacts with the outward conformation of band 3, did not react with HPG-modified membranes. Furthermore, we expressed a band 3 mutant in which Arg 901 was replaced by alanine (R901A) on yeast membranes. The kinetic parameters indicated that the R901A mutation affected the rate of conformational change of the band 3 protein. From these results, we conclude that the most C-terminal arginine, Arg 901, has a functional role in the conformational change that is necessary for anion transport.
- Subjects
ARGININE; PROTEINS; ERYTHROCYTES; PYRUVATE kinase; YEAST
- Publication
Journal of Biochemistry, 2006, Vol 139, Issue 5, p903
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvj097