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15 N and 13 C Group‐Selective Techniques Extend the Scope of STD NMR Detection of Weak Host–Guest Interactions and Ligand Screening
Author(s) -
Kövér Katalin E.,
Wéber Edit,
Martinek Tamás A.,
Monostori Éva,
Batta Gyula
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201000317
Subject(s) - biomolecule , ligand (biochemistry) , chemistry , epitope , nuclear magnetic resonance , biology , biochemistry , physics , receptor , antigen , genetics
Abstract Saturation transfer difference (STD) is a valuable tool for studying the binding of small molecules to large biomolecules and for obtaining detailed information on the binding epitopes. Here, we demonstrate that the proposed 15 N/ 13 C variants of group‐selective, “GS‐STD” experiments provide a powerful approach to mapping the binding epitope of a ligand even in the absence of efficient spin diffusion within the target protein. Therefore, these experimental variants broaden the scope of STD studies to smaller and/or more‐dynamic targets. The STD spectra obtained in four different experimental setups (selective 1 H STD, 15 N GS‐STD, 13 C Ar and 13 C aliphatic GS‐STD approaches) revealed that the signal‐intensity pattern of the difference spectra is affected by both the type and the spatial distribution of the excited “transmitter” atoms, as well as by the efficiency of the spin‐diffusion‐mediated magnetization transfer. The performance of the experiments is demonstrated on a system by using the lectin, galectin‐1 and its carbohydrate ligand, lactose.