4.5 Article

ATP dependent charge movement in ATP7B Cu+-ATPase is demonstrated by pre-steady state electrical measurements

期刊

FEBS LETTERS
卷 584, 期 22, 页码 4619-4622

出版社

WILEY
DOI: 10.1016/j.febslet.2010.10.029

关键词

Charge transfer measurement; Copper displacement; Heterologous expression; Phosphoenzyme intermediate; Recombinant Cu+-ATPase; Solid supported membrane

资金

  1. U.S. National Institutes of Health from the NHBLI [RO301-69830]
  2. Italian Ministry of Education, University and Research
  3. Ente Cassa di Risparmio di Firenze
  4. Italian Ministry of Foreign Affairs

向作者/读者索取更多资源

ATP7B is a copper dependent P-type ATPase, required for copper homeostasis. Taking advantage of high yield heterologous expression of recombinant protein, we investigated charge transfer in ATP7B. We detected charge displacement within a single catalytic cycle upon ATP addition and formation of phosphoenzyme intermediate. We attribute this charge displacement to movement of bound copper within ATP7B. Based on specific mutations, we demonstrate that enzyme activation by copper requires occupancy of a site in the N-terminus extension which is not present in other transport ATPases, as well as of a transmembrane site corresponding to the cation binding site of other ATPases. (C) 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

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