Phosphoenzyme formation by purified, reconstituted copper ATPase of Enterococcus hirae

FEBS Letters

Wyler-Duda, Petra¹  Solioz, Marc¹ 

[1] Institute of Clinical Pharmacology, Murtenstrasse 35, University of Berne, 3010 Berne, Switzerland

关键词: Copper;    P-type ATPase;    Purification;    Phosphorylation;    Vanadate;    (Enterococcus hirae);    MES;    N-morpholinoethanesulfonic acid;    CHAPS;    3[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate;    C8E5−7;    hexaethylene glycol mono-n-(pentyl;    hexyl;    heptyl) octyl ether;    Zwittergent 3–10;    n-decyl-N;    N-dimethyl-3-ammonio-1-propenesulfonate;    BCDS;    bathocuproine disulfonate;   

DOI  :  10.1016/S0014-5793(96)01306-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Enterococcus hirae CopB ATPase serves in the secretion of excess copper from cells and belongs to the recently discovered, new class of heavy metal transport ATPases. We here report the affinity purification of CopB to near homogeneity and its reconstitution into phospholipid vesicles. In these proteoliposomes, the ATPase formed an acylphosphate reaction intermediate with the γ-phosphate of ATP. ATPase activity and phosphoenzyme formation were inhibited by vanadate with an I ₅₀ of 0.1 mM. Our results suggest that heavy metal and non-heavy metal ATPases operate by the same underlying mechanism.

【 授权许可】

Unknown