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PMID: 16150713 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Structural role of countertransport revealed in Ca(2+) pump crystal structure in the absence of Ca(2+).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 41 ·2005-10-11 ·Pages 14489-96

Obara K, Miyashita N, Xu C, Toyoshima I, Sugita Y, Inesi G, Toyoshima C

Abstract

Ca(2+)-ATPase of sarcoplasmic reticulum is an ATP-powered Ca(2+) pump but also a H(+) pump in the opposite direction with no demonstrated functional role. Here, we report a 2.4-A-resolution crystal structure of the Ca(2+)-ATPase in the absence of Ca(2+) stabilized by two inhibitors, dibutyldihydroxybenzene, which bridges two transmembrane helices, and thapsigargin, also bound in the membrane region. Now visualized are water and several phospholipid molecules, one of which occupies a cleft between two transmembrane helices. Atomic models of the Ca(2+) binding sites with explicit hydrogens derived by continuum electrostatic calculations show how water and protons fill the space and compensate charge imbalance created by Ca(2+)-release. They suggest that H(+) countertransport is a consequence of a requirement for maintaining structural integrity of the empty Ca(2+)-binding sites. For this reason, cation countertransport is probably mandatory for all P-type ATPases and possibly accompanies transport of water as well.

MeSH Terms
Animals Biological Transport, Active/physiology Biophysical Phenomena Biophysics COS Cells Calcium-Transporting ATPases/chemistry,genetics,metabolism,ultrastructure Chickens Chlorocebus aethiops Cloning, Molecular Crystallography Ion Transport/physiology Models, Molecular Mutagenesis, Site-Directed Phenol Protons Sarcoplasmic Reticulum Calcium-Transporting ATPases Static Electricity Thapsigargin Water/chemistry
Chemicals
Protons Water Phenol Thapsigargin Sarcoplasmic Reticulum Calcium-Transporting ATPases Calcium-Transporting ATPases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Obara Koji
Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.
Miyashita Naoyuki
Xu Cheng
Toyoshima Itaru
Sugita Yuji
Inesi Giuseppe
Toyoshima Chikashi
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0506222102
Published
2005-10-11
Epub
2005-00-06
Pages
14489-96
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1253571
Subset
IM
Databases
PDB
2AGV
External Links
PubMed source DOI: 10.1073/pnas.0506222102 PMC Full Text View journal details
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