Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
48
pubmed:dateCreated
2007-11-29
pubmed:abstractText
The analytical toolkit developed for investigations into water-soluble protein folding has yet to be applied in earnest to membrane proteins. A major problem is the difficulty in collecting kinetic data, which are crucial to understanding any reaction. Here, we combine kinetic and thermodynamic studies of the reversible unfolding of an alpha-helical membrane protein to provide a definitive value for the reaction free energy and a means to probe the transition state. Our analyses show that the major unfolding step in the SDS-induced denaturation of bacteriorhodopsin involves a reduction in alpha-helical structure and proceeds with a large free-energy change; both our equilibrium and kinetic measurements predict that the free energy of unfolding in the absence of denaturant is +20 kcal.mol(-1), with an associated m-value of 25 kcal.mol(-1). The rate of unfolding in the absence of denaturant, k(u)(H(2)O), is surprisingly very slow ( approximately 10(-15) s(-1)). The kinetics also give information on the transition state for this major unfolding step, with a value for beta (m(f)/[m(f) + m(u)]) of approximately 0.1, indicating that the transition state is close to the unfolded state. We thus present a basis for mapping the structural and energetic properties of membrane protein folding by mutagenesis and classical kinetics.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-10563824, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-10860735, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-10984586, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-11798093, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-11960728, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-122264, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-1318849, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-14659758, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-15215473, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-15383660, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-16023672, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-16593536, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-1849896, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-1967832, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-2006195, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-2605250, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-2748571, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-3434802, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-3717552, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-3756303, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-4418026, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-4940442, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-6514811, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-6691969, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-7085614, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-7217055, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-7407071, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-7749918, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-8356089, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-8376389, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-8639552, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-870032, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-8889171, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-891952, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-8993333, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-9153430, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-9219525, http://linkedlifedata.com/resource/pubmed/commentcorrection/18025476-9710577
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
1091-6490
pubmed:author
pubmed:issnType
Electronic
pubmed:day
27
pubmed:volume
104
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
18970-5
pubmed:dateRevised
2010-9-15
pubmed:meshHeading
pubmed:year
2007
pubmed:articleTitle
Combined kinetic and thermodynamic analysis of alpha-helical membrane protein unfolding.
pubmed:affiliation
Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol BS8 1TD, United Kingdom. p.curnow@bristol.ac.uk
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't