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Functional dynamics of human FKBP12 revealed by methyl C-13 rotating frame relaxation dispersion NMR spectroscopy

Brath, Ulrika LU ; Akke, Mikael LU ; Yang, DW; Kay, LE and Mulder, Frans LU (2006) In Journal of the American Chemical Society 128(17). p.5718-5727
Abstract
Transverse relaxation dispersion NMR spectroscopy can provide atom-specific information about time scales, populations, and the extent of structural reorganization in proteins under equilibrium conditions. A method is described that uses side-chain methyl groups as local reporters for conformational transitions taking place in the microsecond regime. The experiment measures carbon nuclear spin relaxation rates in the presence of continuous wave off-resonance irradiation, in proteins uniformly enriched with C-13, and partially randomly labeled with 2 H. The method was applied to human FK-506 binding protein (FKBP12), which uses a common surface for binding substrates in its dual role as both an immunophilin and folding assistant.... (More)
Transverse relaxation dispersion NMR spectroscopy can provide atom-specific information about time scales, populations, and the extent of structural reorganization in proteins under equilibrium conditions. A method is described that uses side-chain methyl groups as local reporters for conformational transitions taking place in the microsecond regime. The experiment measures carbon nuclear spin relaxation rates in the presence of continuous wave off-resonance irradiation, in proteins uniformly enriched with C-13, and partially randomly labeled with 2 H. The method was applied to human FK-506 binding protein (FKBP12), which uses a common surface for binding substrates in its dual role as both an immunophilin and folding assistant. Conformational dynamics on a time scale of similar to 130 mu s were detected for methyl groups located in the substrate binding pocket, demonstrating its plasticity in the absence of substrate. The spatial arrangement of affected side-chain atoms suggests that substrate recognition involves the rapid relative movement of the subdomain comprising residues Ala81-Thr96 and that the observed dynamics play an important role in facilitating the interaction of this protein with its many partners, including calcineurin. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of the American Chemical Society
volume
128
issue
17
pages
5718 - 5727
publisher
The American Chemical Society
external identifiers
  • wos:000237389900042
  • scopus:33646557015
ISSN
1520-5126
DOI
10.1021/ja0570279
language
English
LU publication?
yes
id
2e590625-1183-4f71-b7e8-fbb2b2c2c245 (old id 410018)
date added to LUP
2007-08-29 09:26:52
date last changed
2019-02-20 07:56:56
@article{2e590625-1183-4f71-b7e8-fbb2b2c2c245,
  abstract     = {Transverse relaxation dispersion NMR spectroscopy can provide atom-specific information about time scales, populations, and the extent of structural reorganization in proteins under equilibrium conditions. A method is described that uses side-chain methyl groups as local reporters for conformational transitions taking place in the microsecond regime. The experiment measures carbon nuclear spin relaxation rates in the presence of continuous wave off-resonance irradiation, in proteins uniformly enriched with C-13, and partially randomly labeled with 2 H. The method was applied to human FK-506 binding protein (FKBP12), which uses a common surface for binding substrates in its dual role as both an immunophilin and folding assistant. Conformational dynamics on a time scale of similar to 130 mu s were detected for methyl groups located in the substrate binding pocket, demonstrating its plasticity in the absence of substrate. The spatial arrangement of affected side-chain atoms suggests that substrate recognition involves the rapid relative movement of the subdomain comprising residues Ala81-Thr96 and that the observed dynamics play an important role in facilitating the interaction of this protein with its many partners, including calcineurin.},
  author       = {Brath, Ulrika and Akke, Mikael and Yang, DW and Kay, LE and Mulder, Frans},
  issn         = {1520-5126},
  language     = {eng},
  number       = {17},
  pages        = {5718--5727},
  publisher    = {The American Chemical Society},
  series       = {Journal of the American Chemical Society},
  title        = {Functional dynamics of human FKBP12 revealed by methyl C-13 rotating frame relaxation dispersion NMR spectroscopy},
  url          = {http://dx.doi.org/10.1021/ja0570279},
  volume       = {128},
  year         = {2006},
}