Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1996-10-17
pubmed:abstractText
The equilibrium unfolding reaction of the C-terminal 80-amino-acid dimeric DNA-binding domain of human papillomavirus (HPV) strain 16 E2 protein has been investigated using fluorescence, far-UV CD, and equilibrium sedimentation. The stability of the HPV-16 E2 DNA-binding domain is concentration-dependent, and the unfolding reaction is well described as a two-state transition from folded dimer to unfolded monomer. The conformational stability of the protein, delta GH2O, was found to be 9.8 kcal/mol at pH 5.6, with the corresponding equilibrium unfolding/dissociation constant, Ku, being 6.5 x 10(-8) M. Equilibrium sedimentation experiments give a Kd of 3.0 x 10(-8) M, showing an excellent agreement between the two different techniques. Denaturation by temperature followed by the change in ellipticity also shows a concomitant disappearance of secondary and tertiary structures. The Ku changes dramatically at physiologically relevant pH's: with a change in pH from 6.1 to 7.0, it goes from 5.5 x 10(-8) M to 4.4 x 10(10) M. Our results suggest that, at the very low concentration of protein where DNA binding is normally measured (e.g., 10(-11) M), the protein is predominantly monomeric and unfolded. They also stress the importance of the coupling between folding and DNA binding.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1316493, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1321680, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1328886, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1390732, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-14268786, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1472508, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1587266, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1655748, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1662805, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1663669, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-1944532, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-2377232, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-2819054, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-3293583, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-3773761, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-4416801, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-6452580, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-7756976, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-7800477, http://linkedlifedata.com/resource/pubmed/commentcorrection/8745409-7849029
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0961-8368
pubmed:author
pubmed:issnType
Print
pubmed:volume
5
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
310-9
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Equilibrium dissociation and unfolding of the dimeric human papillomavirus strain-16 E2 DNA-binding domain.
pubmed:affiliation
Department of Chemistry, University of Cambridge, United Kingdom.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't