Fast Structure-Based Assignment of 15N HSQC Spectra of Selectively 15N-Labeled Paramagnetic Proteins

Pintacuda, Guido, Keniry, Max A., Huber, Thomas, Park, Ah Young, Dixon, Nicholas E. and Otting, Gottfried (2004) Fast Structure-Based Assignment of 15N HSQC Spectra of Selectively 15N-Labeled Paramagnetic Proteins. Journal of the American Chemical Society, 126 9: 2963-2970. doi:10.1021/ja039339m


Author Pintacuda, Guido
Keniry, Max A.
Huber, Thomas
Park, Ah Young
Dixon, Nicholas E.
Otting, Gottfried
Title Fast Structure-Based Assignment of 15N HSQC Spectra of Selectively 15N-Labeled Paramagnetic Proteins
Formatted title
Fast Structure-Based Assignment of 15N HSQC Spectra of Selectively 15N-Labeled Paramagnetic Proteins
Journal name Journal of the American Chemical Society   Check publisher's open access policy
ISSN 0002-7863
Publication date 2004-02-13
Sub-type Article (original research)
DOI 10.1021/ja039339m
Volume 126
Issue 9
Start page 2963
End page 2970
Total pages 8
Editor P. Stang
Place of publication Washington D. C.
Publisher American Chemical Society
Language eng
Subject 250303 Physical Organic Chemistry
250699 Theoretical and Computational Chemistry not elsewhere classified
250502 Physical Chemistry of Macromolecules
250503 Characterisation of Macromolecules
Abstract A novel strategy for fast NMR resonance assignment of N-15 HSQC spectra of proteins is presented. It requires the structure coordinates of the protein, a paramagnetic center, and one or more residue-selectively N-15-labeled samples. Comparison of sensitive undecoupled N-15 HSQC spectra recorded of paramagnetic and diamagnetic samples yields data for every cross-peak on pseudocontact shift, paramagnetic relaxation enhancement, cross-correlation between Curie-spin and dipole-dipole relaxation, and residual dipolar coupling. Comparison of these four different paramagnetic quantities with predictions from the three-dimensional structure simultaneously yields the resonance assignment and the anisotropy of the susceptibility tensor of the paramagnetic center. The method is demonstrated with the 30 kDa complex between the N-terminal domain of the epsilon subunit and the theta subunit of Escherichia Coll DNA polymerase III. The program PLATYPUS was developed to perform the assignment, provide a measure of reliability of the assignment, and determine the susceptibility tensor anisotropy.
Keyword NMR resonance
DNA-polymerase-III
Chemistry, Multidisciplinary
Residual Dipolar Couplings
Escherichia-coli
Pseudocontact Shifts
Cross-correlation
Molecular-dynamics
Alignment Tensors
Nmr-spectroscopy
Epsilon-subunit
Q-Index Code C1

 
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Created: Tue, 05 Jun 2007, 11:45:52 EST by Mrs Leith Woodall on behalf of School of Chemistry & Molecular Biosciences