Linked Life Data

20627162

Source:http://linkedlifedata.com/resource/pubmed/id/20627162

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2010-7-14
pubmed:abstractText
F(1)-ATPase (F(1)) is the water-soluble portion of ATP synthase and a rotary molecular motor in which the rotary shaft, the gamma subunit, rotates with 120 degrees steps against the alpha(3)beta(3) stator ring upon ATP hydrolysis. While the crystal structures of F(1) exhibit essentially one stable conformational state of F(1), single-molecule rotation studies revealed that there are two stable conformations of F(1) in each 120 degrees step: the ATP-binding dwell state and the catalytic dwell state. This chapter provides the experimental procedure for the determination of which catalytic state the crystal structures of F(1) represent, by the use of a cross-linking technique in the single-molecule rotation assay. The beta and gamma subunits are cross-linked through a disulfide bond between two cysteine residues genetically introduced at the positions where the beta and gamma subunits have a specific contact in the crystal structures of the ADP-bound form. In the single-molecule rotation assay, the cross-linked F(1) shows a pause at the catalytic dwell state that corresponds to the dwell angle in one turn where the beta subunit undergoes ATP hydrolysis. Thus, this experiment reveals not only that the crystal structure represents the catalytic dwell state but also that the ADP-bound beta subunit represents the catalytically active state. A protocol for inhibition of the wild-type F(1) with chemical inhibitors such as adenosine-5'-(beta,gamma-imino)-triphosphate (AMP-PNP) or/and N(3)(-) under crystallization conditions is also provided.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1557-7988
pubmed:author
pubmed:copyrightInfo
Copyright (c) 2010 Elsevier Inc. All rights reserved.
pubmed:issnType
Electronic
pubmed:volume
475
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
279-96
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Measurement of the conformational state of F(1)-ATPase by single-molecule rotation.
pubmed:affiliation
The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't