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Distance mapping of protein-binding sites using spin-labeled oligosaccharide ligands

Jain, Nitin U.; Venot, Andre; Umemoto, Kimiko; Leffler, Hakon LU and Prestegard, James H. (2001) In Protein Science 10(11). p.2393-2400
Abstract
The binding of a nitroxide spin-labeled analog of N-acetyllactosamine to galectin-3, a mammalian lectin of 26 kD size, is studied to map the binding sites of this small oligosaccharide on the protein surface. Perturbation of intensities of cross-peaks in the (15)N heteronuclear single quantum coherence (HSQC) spectrum of full-length galectin-3 owing to the bound spin label is used qualitatively to identify protein residues proximate to the binding site for N-acetyllactosamine. A protocol for converting intensity measurements to a more quantitative determination of distances between discrete protein amide protons and the bound spin label is then described. This protocol is discussed as part of a drug design strategy in which subsequent... (More)
The binding of a nitroxide spin-labeled analog of N-acetyllactosamine to galectin-3, a mammalian lectin of 26 kD size, is studied to map the binding sites of this small oligosaccharide on the protein surface. Perturbation of intensities of cross-peaks in the (15)N heteronuclear single quantum coherence (HSQC) spectrum of full-length galectin-3 owing to the bound spin label is used qualitatively to identify protein residues proximate to the binding site for N-acetyllactosamine. A protocol for converting intensity measurements to a more quantitative determination of distances between discrete protein amide protons and the bound spin label is then described. This protocol is discussed as part of a drug design strategy in which subsequent perturbation of chemical shifts of distance mapped amide cross-peaks can be used effectively to screen a library of compounds for other ligands that bind to the target protein at distances suitable for chemical linkage to the primary ligand. This approach is novel in that it bypasses the need for structure determination and resonance assignment of the target protein. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Nitroxide spin label, galectin-3, drug design, drug screening, distance mapping
in
Protein Science
volume
10
issue
11
pages
2393 - 2400
publisher
The Protein Society
external identifiers
  • pmid:11604544
  • scopus:0034775555
ISSN
1469-896X
DOI
10.1110/ps.17401
language
English
LU publication?
yes
id
ab0922fe-6c5a-4c75-86d4-bf72f96bd228 (old id 1123032)
date added to LUP
2008-06-30 16:14:33
date last changed
2018-01-07 06:04:40
@article{ab0922fe-6c5a-4c75-86d4-bf72f96bd228,
  abstract     = {The binding of a nitroxide spin-labeled analog of N-acetyllactosamine to galectin-3, a mammalian lectin of 26 kD size, is studied to map the binding sites of this small oligosaccharide on the protein surface. Perturbation of intensities of cross-peaks in the (15)N heteronuclear single quantum coherence (HSQC) spectrum of full-length galectin-3 owing to the bound spin label is used qualitatively to identify protein residues proximate to the binding site for N-acetyllactosamine. A protocol for converting intensity measurements to a more quantitative determination of distances between discrete protein amide protons and the bound spin label is then described. This protocol is discussed as part of a drug design strategy in which subsequent perturbation of chemical shifts of distance mapped amide cross-peaks can be used effectively to screen a library of compounds for other ligands that bind to the target protein at distances suitable for chemical linkage to the primary ligand. This approach is novel in that it bypasses the need for structure determination and resonance assignment of the target protein.},
  author       = {Jain, Nitin U. and Venot, Andre and Umemoto, Kimiko and Leffler, Hakon and Prestegard, James H.},
  issn         = {1469-896X},
  keyword      = {Nitroxide spin label,galectin-3,drug design,drug screening,distance mapping},
  language     = {eng},
  number       = {11},
  pages        = {2393--2400},
  publisher    = {The Protein Society},
  series       = {Protein Science},
  title        = {Distance mapping of protein-binding sites using spin-labeled oligosaccharide ligands},
  url          = {http://dx.doi.org/10.1110/ps.17401},
  volume       = {10},
  year         = {2001},
}