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Rapid and accurate structure determination of coiled‐coil domains using NMR dipolar couplings: Application to cGMP‐dependent protein kinase Iα
Author(s) -
Schnell Jason R.,
Zhou GuoPing,
Zweckstetter Markus,
Rigby Alan C.,
Chou James J.
Publication year - 2005
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.051528905
Subject(s) - coiled coil , electromagnetic coil , protein structure , dipole , residual dipolar coupling , nuclear magnetic resonance spectroscopy , relaxation (psychology) , crystallography , chemistry , biophysics , nuclear magnetic resonance , physics , biology , organic chemistry , quantum mechanics , neuroscience
Coiled‐coil motifs play essential roles in protein assembly and molecular recognition, and are therefore the targets of many ongoing structural and functional studies. However, owing to the dynamic nature of many of the smaller coiled‐coil domains, crystallization for X‐ray studies is very challenging. Determination of elongated structures using standard NMR approaches is inefficient and usually yields low‐resolution structures due to accumulation of small errors over long distances. Here we describe a solution NMR approach based on residual dipolar couplings (RDCs) for rapid and accurate structure determination of coiled‐coil dimers. Using this approach, we were able to determine the high‐resolution structure of the coiled‐coil domain of cGMP‐dependent protein kinase Iα, a protein of previously unknown structure that is critical for physiological relaxation of vascular smooth muscle. This approach can be extended to solve coiled‐coil structures with higher order assemblies.