The ATP-binding site of Ca2+-ATPase revealed by electron image analysis

Koji Yonekura; David L. Stokes; Hiroyuki Sasabe; Chikashi Toyoshima

doi:10.1016/S0006-3495(97)78752-6

The ATP-binding site of Ca²⁺-ATPase revealed by electron image analysis

Koji Yonekura, David L. Stokes, Hiroyuki Sasabe, Chikashi Toyoshima

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The location of the ATP-binding site of a P-type ion pump, Ca²⁺- ATPase from rabbit sarcoplasmic reticulum, was examined by cryoelectron microscopy. A nonhydrolyzable analog of ATP, β,γ-bidentate chromium (III) complex of ATP (CrATP), was used to stabilize the enzyme in the Ca²⁺- occluded state. Tubular crystals were then induced by vanadate in the presence of EGTA, keeping CrATP bound to the enzyme. The three-dimensional structures of the crystals were determined at 14 Å resolution by cryoelectron microscopy and helical image analysis. Statistical comparison of the structures with and without CrATP showed clear and significant differences at the groove proposed previously as the ATP-binding pocket.

Original language	English
Pages (from-to)	997-1005
Number of pages	9
Journal	Biophysical Journal
Volume	72
Issue number	3
DOIs	https://doi.org/10.1016/S0006-3495(97)78752-6
Publication status	Published - 1997 Mar
Externally published	Yes

ASJC Scopus subject areas

Biophysics

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title = "The ATP-binding site of Ca2+-ATPase revealed by electron image analysis",

abstract = "The location of the ATP-binding site of a P-type ion pump, Ca2+- ATPase from rabbit sarcoplasmic reticulum, was examined by cryoelectron microscopy. A nonhydrolyzable analog of ATP, β,γ-bidentate chromium (III) complex of ATP (CrATP), was used to stabilize the enzyme in the Ca2+- occluded state. Tubular crystals were then induced by vanadate in the presence of EGTA, keeping CrATP bound to the enzyme. The three-dimensional structures of the crystals were determined at 14 {\AA} resolution by cryoelectron microscopy and helical image analysis. Statistical comparison of the structures with and without CrATP showed clear and significant differences at the groove proposed previously as the ATP-binding pocket.",

author = "Koji Yonekura and Stokes, {David L.} and Hiroyuki Sasabe and Chikashi Toyoshima",

note = "Funding Information: We gratefully thank Prof. T. Tsukihara of the Institute for Protein Re-search, Osaka University, for a set of real-space-fitting programs and K. Tani for implementing these programs on our computers. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.",

year = "1997",

month = mar,

doi = "10.1016/S0006-3495(97)78752-6",

language = "English",

volume = "72",

pages = "997--1005",

journal = "Biophysical Journal",

issn = "0006-3495",

publisher = "Biophysical Society",

number = "3",

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T1 - The ATP-binding site of Ca2+-ATPase revealed by electron image analysis

AU - Yonekura, Koji

AU - Stokes, David L.

AU - Sasabe, Hiroyuki

AU - Toyoshima, Chikashi

N1 - Funding Information: We gratefully thank Prof. T. Tsukihara of the Institute for Protein Re-search, Osaka University, for a set of real-space-fitting programs and K. Tani for implementing these programs on our computers. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.

PY - 1997/3

Y1 - 1997/3

N2 - The location of the ATP-binding site of a P-type ion pump, Ca2+- ATPase from rabbit sarcoplasmic reticulum, was examined by cryoelectron microscopy. A nonhydrolyzable analog of ATP, β,γ-bidentate chromium (III) complex of ATP (CrATP), was used to stabilize the enzyme in the Ca2+- occluded state. Tubular crystals were then induced by vanadate in the presence of EGTA, keeping CrATP bound to the enzyme. The three-dimensional structures of the crystals were determined at 14 Å resolution by cryoelectron microscopy and helical image analysis. Statistical comparison of the structures with and without CrATP showed clear and significant differences at the groove proposed previously as the ATP-binding pocket.

AB - The location of the ATP-binding site of a P-type ion pump, Ca2+- ATPase from rabbit sarcoplasmic reticulum, was examined by cryoelectron microscopy. A nonhydrolyzable analog of ATP, β,γ-bidentate chromium (III) complex of ATP (CrATP), was used to stabilize the enzyme in the Ca2+- occluded state. Tubular crystals were then induced by vanadate in the presence of EGTA, keeping CrATP bound to the enzyme. The three-dimensional structures of the crystals were determined at 14 Å resolution by cryoelectron microscopy and helical image analysis. Statistical comparison of the structures with and without CrATP showed clear and significant differences at the groove proposed previously as the ATP-binding pocket.

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JO - Biophysical Journal

JF - Biophysical Journal

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ER -

The ATP-binding site of Ca²⁺-ATPase revealed by electron image analysis

Abstract

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