期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 322, 期 2, 页码 425-440出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/S0022-2836(02)00739-8
关键词
hydrophobic cavities; ligand-protein interactions; xenon binding; multiple isomorphous replacement; protein conformation assay
Xenon-binding sites in proteins have led to a number of applications of xenon in biochemical and structural studies. Here we further develop the utility of Xe-129 NMR in characterizing specific xenon-protein interactions. The sensitivity of the Xe-129 chemical shift to its local environment and the intense signals attainable by optical pumping make xenon a useful NMR reporter of its own interactions with proteins. A method for detecting specific xenon-binding interactions by analysis of Xe-129 chemical shift data is illustrated using the maltose binding protein (MBP) from Escherichia coli as an example. The crystal structure of MBP in the presence of 8 atm of xenon confirms the binding site determined from NMR data. Changes in the structure of the xenon-binding cavity upon the binding of maltose by the protein can account for the sensitivity of the Xe-129 chemical shift to MBP conformation. Xe-129 NMR data for xenon in solution with a number of cavity containing phage T4 lysozyme mutants show that xenon can report on cavity structure. In particular, a correlation exists between cavity size and the binding-induced Xe-129 chemical shift. Further applications of Xe-129 NMR to biochemical assays, including the screening of proteins for xenon binding for crystallography are considered. (C) 2002 Elsevier Science Ltd. All rights reserved
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