Rescuing a destabilized protein fold through backbone cyclization.
J Mol Biol
; 308(5): 1045-62, 2001 May 18.
Article
en En
| MEDLINE
| ID: mdl-11352590
ABSTRACT
We describe the physicochemical characterization of various circular and linear forms of the approximately 60 residue N-terminal Src homology 3 (SH3) domain from the murine c-Crk adapter protein. Structural, dynamic, thermodynamic, kinetic and biochemical studies reveal that backbone circularization does not prevent the adoption of the natural folded structure in any of the circular proteins. Both the folding and unfolding rate of the protein increased slightly upon circularization. Circularization did not lead to a significant thermodynamic stabilization of the full-length protein, suggesting that destabilizing enthalpic effects (e.g. strain) negate the expected favorable entropic contribution to overall stability. In contrast, we find circularization results in a dramatic stabilization of a truncated version of the SH3 domain lacking a key glutamate residue. The ability to rescue the destabilized mutant indicates that circularization may be a useful tool in protein engineering programs geared towards generating minimized proteins.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Ingeniería de Proteínas
/
Proteínas Proto-Oncogénicas
/
Pliegue de Proteína
/
Dominios Homologos src
Límite:
Animals
Idioma:
En
Revista:
J Mol Biol
Año:
2001
Tipo del documento:
Article
País de afiliación:
Estados Unidos