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

Protein Science : a Publication of the Protein Society
N U JainJames H Prestegard

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.

References

Jan 1, 1989·Methods in Enzymology·P A Kosen
May 1, 1994·Journal of Magnetic Resonance. Series B·L YuS W Fesik
Nov 29, 1996·Science·S B ShukerS W Fesik
May 28, 1998·The Journal of Biological Chemistry·J SeetharamanJ M Rini
Oct 16, 1999·Glycobiology·D N Cooper, S H Barondes
Dec 2, 1999·Biochimica Et Biophysica Acta·R C Hughes
Mar 16, 2000·Molecular and Biochemical Parasitology·C J GreenhalghS E Newton
Oct 12, 2000·Clinical Immunology : the Official Journal of the Clinical Immunology Society·F T Liu
Feb 24, 2001·Results and Problems in Cell Differentiation·H Leffler
Jan 5, 2005·Journal of Biomolecular NMR·Giuseppe NicastroAnnalisa Pastore

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Citations

Jun 19, 2008·Journal of Biomolecular NMR·Hanna E LindforsMarcellus Ubbink
Oct 22, 2003·Current Opinion in Structural Biology·Heide KogelbergJesús Jiménez-Barbero
Feb 12, 2009·The Journal of Biological Chemistry·Sagaya Theresa Leena PhilominathanJoshua Sakon
Jan 12, 2013·Current Protein & Peptide Science·Maria del Carmen Fernández-AlonsoJesús Jiménez-Barbero
Apr 18, 2018·Chemical Communications : Chem Comm·Ana Ardá, Jesús Jiménez-Barbero
May 22, 2008·Molecular BioSystems·Emiliano GemmaAlison N Hulme
Apr 13, 2006·Proteins·Yaroslav Ryabov, David Fushman
Feb 25, 2015·Quarterly Reviews of Biophysics·Nicholas J Anthis, G Marius Clore
Jan 30, 2009·Protein Science : a Publication of the Protein Society·Richard C PageTimothy A Cross
Apr 28, 2007·Chemistry & Biology·Shan LiuJames H Prestegard

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