Skip navigation
Skip navigation

DOTA-Amide Lanthanide Tag for Reliable Generation of Pseudocontact Shifts in Protein NMR Spectra

Graham, Bim; Loh, Choy; Swarbrick, James; Ung, Phuc; Shin, James; Yagi, Hiromasa; Jia, Xinying; Chhabra, Sandeep; Barlow, Nicholas; Pintacuda, Guido; Otting, Gottfried; Huber, Thomas

Description

Structural studies of proteins and protein-ligand complexes by nuclear magnetic resonance (NMR) spectroscopy can be greatly enhanced by site-specific attachment of lanthanide ions to create paramagnetic centers. In particular, pseudocontact shifts (PCS) generated by paramagnetic lanthanides contain important and unique long-range structure information. Here, we present a high-affinity lanthanide binding tag that can be attached to single cysteine residues of proteins. The new tag has many...[Show more]

dc.contributor.authorGraham, Bim
dc.contributor.authorLoh, Choy
dc.contributor.authorSwarbrick, James
dc.contributor.authorUng, Phuc
dc.contributor.authorShin, James
dc.contributor.authorYagi, Hiromasa
dc.contributor.authorJia, Xinying
dc.contributor.authorChhabra, Sandeep
dc.contributor.authorBarlow, Nicholas
dc.contributor.authorPintacuda, Guido
dc.contributor.authorOtting, Gottfried
dc.contributor.authorHuber, Thomas
dc.date.accessioned2015-12-10T22:56:51Z
dc.identifier.issn1043-1802
dc.identifier.urihttp://hdl.handle.net/1885/60409
dc.description.abstractStructural studies of proteins and protein-ligand complexes by nuclear magnetic resonance (NMR) spectroscopy can be greatly enhanced by site-specific attachment of lanthanide ions to create paramagnetic centers. In particular, pseudocontact shifts (PCS) generated by paramagnetic lanthanides contain important and unique long-range structure information. Here, we present a high-affinity lanthanide binding tag that can be attached to single cysteine residues of proteins. The new tag has many advantageous features that are not available in this combination from previously published tags: (i) it binds lanthanide ions very tightly, minimizing the generation of nonspecific effects, (ii) it produces PCSs with high reliability as its bulkiness prevents complete motional averaging of PCSs, (iii) it can be attached to single cysteine residues, alleviating the need of detailed prior knowledge of the 3D structure of the target protein, and (iv) it does not display conformational exchange phenomena that would increase the number of signals in the NMR spectrum. The performance of the tag is demonstrated with the N-terminal domain of the E. coli arginine repressor and the A28C mutant of human ubiquitin.
dc.publisherAmerican Chemical Society
dc.sourceBioconjugate Chemistry
dc.subjectKeywords: 1,4,7,10 tetraazacyclododecane 1,4,7,10 tetraacetic acid; 2 bromo n (1 phenylethyl)acetamide; 2 chloro n [2 (pyridin 2 yldisulfanyl)ethyl]acetamide; 2,2',2'' (1,4,7,10 tetraazacyclododecane 1,4,7 triyl)tris[n (1 phenylethyl)acetamide]; 2,2',2'' [10 [2 oxo
dc.titleDOTA-Amide Lanthanide Tag for Reliable Generation of Pseudocontact Shifts in Protein NMR Spectra
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume22
dc.date.issued2011
local.identifier.absfor030606 - Structural Chemistry and Spectroscopy
local.identifier.absfor060112 - Structural Biology (incl. Macromolecular Modelling)
local.identifier.absfor030503 - Organic Chemical Synthesis
local.identifier.ariespublicationu4005981xPUB538
local.type.statusPublished Version
local.contributor.affiliationGraham, Bim, Monash University
local.contributor.affiliationLoh, Choy, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationSwarbrick, James, Monash University
local.contributor.affiliationUng, Phuc, Monash Institute of Pharmaceutical Sciences
local.contributor.affiliationShin, James, Monash Institute of Pharmaceutical Sciences
local.contributor.affiliationYagi, Hiromasa, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationJia, Xinying, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationChhabra, Sandeep, Monash Institute of Pharmaceutical Sciences
local.contributor.affiliationBarlow, Nicholas, Monash Institute of Pharmaceutical Sciences
local.contributor.affiliationPintacuda, Guido, Ecole Normale Superieure de Lyon
local.contributor.affiliationHuber, Thomas, 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.startpage2118
local.bibliographicCitation.lastpage2125
local.identifier.doi10.1021/bc200353c
local.identifier.absseo860803 - Human Pharmaceutical Treatments (e.g. Antibiotics)
dc.date.updated2016-02-24T10:24:35Z
local.identifier.scopusID2-s2.0-80054814367
local.identifier.thomsonID000295915100027
CollectionsANU Research Publications

Download

File Description SizeFormat Image
01_Graham_DOTA-Amide_Lanthanide_Tag_for_2011.pdf1.68 MBAdobe PDF    Request a copy


Items in Open Research are protected by copyright, with all rights reserved, unless otherwise indicated.

Updated:  19 May 2020/ Responsible Officer:  University Librarian/ Page Contact:  Library Systems & Web Coordinator