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Measurement of dissociation constants of high-molecular weight protein-protein complexes by transferred 15 N-relaxation

Su, Xun-Cheng; Jergic, Slobodan; Ozawa, Kiyoshi; Burns, Nicolas; Dixon, Nicholas; Otting, Gottfried

Description

The use of 15N-relaxation data for determination of the dissociation constant of a protein-protein complex is proposed for the situation where a 15N-labeled protein is bound to an unlabeled protein of high molecular weight, and the chemical exchange between bound and free protein is fast on the NMR time scale. The approach is shown to be suitable for estimating dissociation constants in the micromolar to millimolar range, using protein solutions at relatively low concentration. An example is...[Show more]

dc.contributor.authorSu, Xun-Cheng
dc.contributor.authorJergic, Slobodan
dc.contributor.authorOzawa, Kiyoshi
dc.contributor.authorBurns, Nicolas
dc.contributor.authorDixon, Nicholas
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2015-12-07T22:54:02Z
dc.identifier.issn0925-2738
dc.identifier.urihttp://hdl.handle.net/1885/27995
dc.description.abstractThe use of 15N-relaxation data for determination of the dissociation constant of a protein-protein complex is proposed for the situation where a 15N-labeled protein is bound to an unlabeled protein of high molecular weight, and the chemical exchange between bound and free protein is fast on the NMR time scale. The approach is shown to be suitable for estimating dissociation constants in the micromolar to millimolar range, using protein solutions at relatively low concentration. An example is shown for the interaction between two subunits from the Escherichia coli DNA polymerase III complex, involving a 15N-labeled fragment of the C-terminal domain of the τ subunit (15 kDa) and the unlabeled α subunit (130 kDa).
dc.publisherKluwer Academic Publishers
dc.sourceJournal of Biomolecular NMR
dc.subjectKeywords: bacterial DNA; DNA directed DNA polymerase gamma; nitrogen 15; protein subunit; article; carboxy terminal sequence; chemical interaction; concentration response; dissociation constant; Escherichia coli; isotope labeling; molecular weight; nitrogen nuclear 15N relaxation; Chemical exchange; Dissociation constant; E. coli DNA polymerase III; Protein-protein complex; Subunits a and t
dc.titleMeasurement of dissociation constants of high-molecular weight protein-protein complexes by transferred 15 N-relaxation
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume38
dc.date.issued2007
local.identifier.absfor069999 - Biological Sciences not elsewhere classified
local.identifier.ariespublicationu4005981xPUB55
local.type.statusPublished Version
local.contributor.affiliationSu, Xun-Cheng, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationJergic, Slobodan, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOzawa, Kiyoshi, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationBurns, Nicolas , College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationDixon, Nicholas, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOtting, Gottfried, College of Physical and Mathematical Sciences, ANU
local.description.embargo2037-12-31
local.bibliographicCitation.issue1
local.bibliographicCitation.startpage65
local.bibliographicCitation.lastpage72
local.identifier.doi10.1007/s10858-007-9147-9
dc.date.updated2015-12-07T12:45:48Z
local.identifier.scopusID2-s2.0-34248547901
CollectionsANU Research Publications

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