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DOTA-M8 : an extremely rigid, high-affinity lanthanide chelating tag for PCS NMR spectroscopy
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 155748
Author(s) Häussinger, Daniel; Huang, Jie-rong; Grzesiek, Stephan
Author(s) at UniBasel Grzesiek, Stephan
Häussinger, Daniel
Year 2009
Title DOTA-M8 : an extremely rigid, high-affinity lanthanide chelating tag for PCS NMR spectroscopy
Journal Journal of the American Chemical Society
Volume 131
Number 41
Pages / Article-Number 14761-7
Keywords paramagnetic NMR, pseudo contact shift, residual dipolar coupling, lanthanide chelating tag, dysprosium, protein
Abstract

A new lanthanide chelating tag (M8) for paramagnetic labeling of biomolecules is presented, which is based on an eight-fold, stereoselectively methyl-substituted DOTA that can be covalently linked to the host molecule by a single disulfide bond. The steric overcrowding of the DOTA scaffold leads to an extremely rigid, kinetically and chemically inert lanthanide chelator. Its steric bulk restricts the motion of the tag relative to the host molecule. These properties result in very large pseudocontact shifts (>5 ppm) and residual dipolar couplings (>20 Hz) for Dy-M8 linked to ubiquitin, which are unprecedented for a small, single-point-attachment tag. Such large pseudocontact shifts should be well detectable even for larger proteins and distances beyond approximately 50 A. Due to its exceptionally high stability and lanthanide affinity M8 can be used under extreme chemical or physical conditions, such as those applied for protein denaturation, or when it is undesirable that buffer or protein react with excess lanthanide ions.

Publisher American Chemical Society
ISSN/ISBN 0002-7863
edoc-URL http://edoc.unibas.ch/dok/A5258750
Full Text on edoc No
Digital Object Identifier DOI 10.1021/ja903233w
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/19785413
ISI-Number WOS:000271271800048
Document type (ISI) Journal Article
 
   

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02/05/2024