Solution structure and folding characteristics of the C-terminal SH3 domain of c-Crk-II.
Crk-II is a signaling adaptor protein that is involved in many cellular processes including apoptosis, proliferation, and differentiation. It has a modular domain architecture consisting of an Src homology 2 domain (SH2) followed by two Src homology 3 (SH3) domains. The structures and ligand-binding properties of the SH2 and the middle SH3 domains are well-characterized. Several studies suggest that the C-terminal SH3 domain plays an important regulatory role in the protein; however, no structural information is available on this domain, and relatively little is known about its binding partners. In the current work, we have solved the solution NMR structure of the C-terminal SH3 domain. The domain adopts the standard SH3 fold comprising a five-stranded beta barrel. In agreement with alignment and modeling studies, the structure indicates that the canonical-binding surface of the SH3 domain is unusually polar and suggests that this domain may not bind typical PXXP ligands or that it may bind them with reduced affinity. Thermodynamic and kinetic studies show that the domain folds in a reversible two-state manner and that the stability of the fold is similar to that observed for other SH3 domains. These studies offer some insight into the likely structural and thermodynamic consequences of point mutations in the cSH3 domain that are known to deregulate Crk-II function. Our results set the stage for a better understanding the role of the cSH3 domain in the context of the full-length protein.
Muralidharan
Vasant
V
Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, New York 10021, USA.
Dutta
Kaushik
K
Cho
Jaehyun
J
Vila-Perello
Miquel
M
Raleigh
Daniel P
DP
Cowburn
David
D
Muir
Tom W
TW
D000595:Amino Acid Sequence; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D050737:Proto-Oncogene Proteins c-crk / Q000737:chemistry*; D016415:Sequence Alignment; D018909:src Homology Domains / Q000502:physiology*
D050737:Proto-Oncogene Proteins c-crk