Advanced

Enhancement of bound-state residual dipolar couplings: Conformational analysis of lactose bound to Galectin-3

Zhuang, TD; Leffler, Hakon LU and Prestegard, JH (2006) In Protein Science 15(7). p.1780-1790
Abstract
Residual dipolar couplings (RDCs) have proven to be a valuable NMR tool that can provide long-range constraints for molecular structure determination. The constraints are orientational in nature and are, thus, highly complementary to conventional distance constraints from NOE data. This complementarity would seem to extend to the study of the geometry of ligands bound to proteins. However, unlike transferred NOEs, where collection, even with a large excess of free ligand, results in measurements dominated by bound contributions, RDCs of exchanging ligands can be dominated by free-state contributions. Here we present a strategy for enhancement of RDCs from bound states that is based on specifically enhancing the alignment of the protein to... (More)
Residual dipolar couplings (RDCs) have proven to be a valuable NMR tool that can provide long-range constraints for molecular structure determination. The constraints are orientational in nature and are, thus, highly complementary to conventional distance constraints from NOE data. This complementarity would seem to extend to the study of the geometry of ligands bound to proteins. However, unlike transferred NOEs, where collection, even with a large excess of free ligand, results in measurements dominated by bound contributions, RDCs of exchanging ligands can be dominated by free-state contributions. Here we present a strategy for enhancement of RDCs from bound states that is based on specifically enhancing the alignment of the protein to which a ligand will bind. The protein is modified by addition of a hydrophobic alkyl tail that anchors it to the bicelles that are a part of the ordering medium needed for RDC measurement. As an illustration, we have added a propyl chain to the C terminus of the carbohydrate recognition domain of the protein, Galectin-3, and report enhanced RDCs that prove consistent with known bound-ligand geometries for this protein. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
transferred NOE, NMR, ligand conformation, rDC, lectin, carbohydrate, binding
in
Protein Science
volume
15
issue
7
pages
1780 - 1790
publisher
The Protein Society
external identifiers
  • wos:000238707200020
  • pmid:16751604
  • scopus:33745698459
ISSN
1469-896X
DOI
10.1110/ps.051994306
language
English
LU publication?
yes
id
73b1ca68-1601-4abf-8668-ba64a1ca7077 (old id 404346)
date added to LUP
2007-10-21 09:20:40
date last changed
2019-02-20 03:38:12
@article{73b1ca68-1601-4abf-8668-ba64a1ca7077,
  abstract     = {Residual dipolar couplings (RDCs) have proven to be a valuable NMR tool that can provide long-range constraints for molecular structure determination. The constraints are orientational in nature and are, thus, highly complementary to conventional distance constraints from NOE data. This complementarity would seem to extend to the study of the geometry of ligands bound to proteins. However, unlike transferred NOEs, where collection, even with a large excess of free ligand, results in measurements dominated by bound contributions, RDCs of exchanging ligands can be dominated by free-state contributions. Here we present a strategy for enhancement of RDCs from bound states that is based on specifically enhancing the alignment of the protein to which a ligand will bind. The protein is modified by addition of a hydrophobic alkyl tail that anchors it to the bicelles that are a part of the ordering medium needed for RDC measurement. As an illustration, we have added a propyl chain to the C terminus of the carbohydrate recognition domain of the protein, Galectin-3, and report enhanced RDCs that prove consistent with known bound-ligand geometries for this protein.},
  author       = {Zhuang, TD and Leffler, Hakon and Prestegard, JH},
  issn         = {1469-896X},
  keyword      = {transferred NOE,NMR,ligand conformation,rDC,lectin,carbohydrate,binding},
  language     = {eng},
  number       = {7},
  pages        = {1780--1790},
  publisher    = {The Protein Society},
  series       = {Protein Science},
  title        = {Enhancement of bound-state residual dipolar couplings: Conformational analysis of lactose bound to Galectin-3},
  url          = {http://dx.doi.org/10.1110/ps.051994306},
  volume       = {15},
  year         = {2006},
}