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Site-Specific Labelling of Proteins with a Rigid Lanthanide-Binding Tag

Su, Xun-Cheng; Dixon, Nicholas; Otting, Gottfried; Huber, Thomas

Description

This paper describes a generic method for the site-specific attachment of lanthanide complexes to proteins through a disulfide bond. The method is demonstrated by the attachment of a lanthanide-binding peptide tag to the single cysteine residue present in the N-terminal DNA-binding domain of the Escherichia coli arginine repressor. Complexes with Y3+, Tb3+, Dy3+, Ho3+, Er3+, Tm3+ and Yb3+ ions were formed and analysed by NMR spectroscopy. Large pseudocontact shifts and residual dipolar...[Show more]

dc.contributor.authorSu, Xun-Cheng
dc.contributor.authorDixon, Nicholas
dc.contributor.authorOtting, Gottfried
dc.contributor.authorHuber, Thomas
dc.date.accessioned2015-12-10T23:01:39Z
dc.identifier.issn1439-4227
dc.identifier.urihttp://hdl.handle.net/1885/61690
dc.description.abstractThis paper describes a generic method for the site-specific attachment of lanthanide complexes to proteins through a disulfide bond. The method is demonstrated by the attachment of a lanthanide-binding peptide tag to the single cysteine residue present in the N-terminal DNA-binding domain of the Escherichia coli arginine repressor. Complexes with Y3+, Tb3+, Dy3+, Ho3+, Er3+, Tm3+ and Yb3+ ions were formed and analysed by NMR spectroscopy. Large pseudocontact shifts and residual dipolar couplings were induced by the lanthanide-binding tag in the protein NMR spectrum, a result indicating that the tag was rigidly attached to the protein. The axial components of the magnetic susceptibility anisotropy tensors determined for the different lanthanide ions were similarly but not identically oriented. A single tag with a single protein attachment site can provide different pseudocontact shifts from different magnetic susceptibility tensors and thus provide valuable nondegenerate long-range structure information in the determination of 3D protein structures by NMR spectroscopy.
dc.publisherWiley-VCH Verlag GMBH
dc.sourceChemBioChem
dc.subjectKeywords: arginine; cysteine; DNA binding protein; lanthanide; anisotropy; article; binding affinity; disulfide bond; DNA binding; Escherichia coli; nuclear magnetic resonance spectroscopy; priority journal; protein analysis; protein binding; protein structure; Ami Lanthanides; NMR spectroscopy; Peptides; Proteins; Structure elucidation
dc.titleSite-Specific Labelling of Proteins with a Rigid Lanthanide-Binding Tag
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume7
dc.date.issued2006
local.identifier.absfor060102 - Bioinformatics
local.identifier.ariespublicationU4217927xPUB631
local.type.statusPublished Version
local.contributor.affiliationSu, Xun-Cheng, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationHuber, Thomas, 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.issue10
local.bibliographicCitation.startpage1599
local.bibliographicCitation.lastpage1604
local.identifier.doi10.1002/cbic.200600142
dc.date.updated2015-12-10T08:29:01Z
local.identifier.scopusID2-s2.0-33749680579
CollectionsANU Research Publications

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