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Amino-acid Type Identification in 15 N-HSQC Spectra by Combinatorial Selective 15 N-labelling

Wu, Peter; Ozawa, Kiyoshi; Jergic, Slobodan; Su, Xun-Cheng; Dixon, Nicholas; Otting, Gottfried

Description

The efficiency of cell-free protein synthesis combined with combinatorial selective 15N-labelling provides a method for the rapid assignment of 15N-HSQC cross-peaks to the 19 different non-proline amino-acid types from five 15N-HSQC spectra. This strategy was explored with two different constructs of the C-terminal domain V of the τ subunit of the Escherichia coli DNA polymerase III holoenzyme, τC16 and τC14. Since each of the five 15N-HSQC spectra contained only about one third of the...[Show more]

dc.contributor.authorWu, Peter
dc.contributor.authorOzawa, Kiyoshi
dc.contributor.authorJergic, Slobodan
dc.contributor.authorSu, Xun-Cheng
dc.contributor.authorDixon, Nicholas
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2015-12-13T23:05:34Z
dc.identifier.issn0925-2738
dc.identifier.urihttp://hdl.handle.net/1885/85600
dc.description.abstractThe efficiency of cell-free protein synthesis combined with combinatorial selective 15N-labelling provides a method for the rapid assignment of 15N-HSQC cross-peaks to the 19 different non-proline amino-acid types from five 15N-HSQC spectra. This strategy was explored with two different constructs of the C-terminal domain V of the τ subunit of the Escherichia coli DNA polymerase III holoenzyme, τC16 and τC14. Since each of the five 15N-HSQC spectra contained only about one third of the cross-peaks present in uniformly labelled samples, spectral overlap was much reduced. All 15N-HSQC cross-peaks of the backbone amides could be assigned to the correct amino-acid type. Availability of the residue-type information greatly assisted the evaluation of the changes in chemical shifts observed for corresponding residues in τC16 vs. those in τC14, and the analysis of the structure and mobility of the C-terminal residues present in τC16 but not in τC14.
dc.publisherKluwer Academic Publishers
dc.sourceJournal of Biomolecular NMR
dc.subjectKeywords: amino acid; DNA directed DNA polymerase gamma; holoenzyme; nitrogen 15; amino acid analysis; article; carboxy terminal sequence; cell free system; controlled study; enzyme subunit; Escherichia coli; isotope labeling; nitrogen nuclear magnetic resonance; n 15N-HSQC; Cell-free protein synthesis; Combinatorial15N-labelling; DNA polymerase III; Resonance assignment; Subunit ?
dc.titleAmino-acid Type Identification in 15 N-HSQC Spectra by Combinatorial Selective 15 N-labelling
dc.typeJournal article
local.description.notesImported from ARIES
local.description.refereedYes
local.identifier.citationvolume34
dc.date.issued2006
local.identifier.absfor060199 - Biochemistry and Cell Biology not elsewhere classified
local.identifier.ariespublicationMigratedxPub14085
local.type.statusPublished Version
local.contributor.affiliationWu, Peter, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOzawa, Kiyoshi, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationJergic, Slobodan, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationSu, Xun-Cheng, 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.startpage13
local.bibliographicCitation.lastpage21
local.identifier.doi10.1007/s10858-005-5021-9
dc.date.updated2015-12-12T08:01:07Z
local.identifier.scopusID2-s2.0-33644517569
CollectionsANU Research Publications

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