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NMR‐Based Screening of Membrane Protein Ligands
Author(s) -
Yanamala Naveena,
Dutta Arpana,
Beck Barbara,
Van Fleet Bart,
Hay Kelly,
Yazbak Ahmad,
Ishima Rieko,
Doemling Alexander,
KleinSeetharaman Judith
Publication year - 2010
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/j.1747-0285.2009.00940.x
Subject(s) - ligand (biochemistry) , nuclear magnetic resonance spectroscopy , chemistry , membrane protein , micelle , drug discovery , protein structure , membrane , biochemistry , stereochemistry , organic chemistry , receptor , aqueous solution
Membrane proteins pose problems for the application of NMR‐based ligand‐screening methods because of the need to maintain the proteins in a membrane mimetic environment such as detergent micelles: they add to the molecular weight of the protein, increase the viscosity of the solution, interact with ligands non‐specifically, overlap with protein signals, modulate protein dynamics and conformational exchange and compromise sensitivity by adding highly intense background signals. In this article, we discuss the special considerations arising from these problems when conducting NMR‐based ligand‐binding studies with membrane proteins. While the use of 13 C and 15 N isotopes is becoming increasingly feasible, 19 F and 1 H NMR‐based approaches are currently the most widely explored. By using suitable NMR parameter selection schemes independent of or exploiting the presence of detergent, 1 H‐based approaches require least effort in sample preparation because of the high sensitivity and natural abundance of 1 H in both, ligand and protein. On the other hand, the 19 F nucleus provides an ideal NMR probe because of its similarly high sensitivity to that of 1 H and the lack of natural 19 F background in biologic systems. Despite its potential, the use of NMR spectroscopy is highly underdeveloped in the area of drug discovery for membrane proteins.