Structural investigation of ribonuclease A conformational preferences using high pressure protein crystallography

Hydrostatic pressure in range 0.1-1.5 GPa is used to modify biological system behaviour mostly in biophysical studies of proteins in solution. Due to specific influence on the system equilibrium high pressure can act as a filter that enables to identify and investigate higher energy protein conforme...

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Detalles Bibliográficos
Autores: Kurpiewska, Katarzyna, Dziubek, Kamil, Katrusiak, Andrzej, Font i Sadurní, Josep, Ribó i Panosa, Marc, Vilanova i Brugués, Maria, Lewiński, Krzysztof
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10256/12346
Acceso en línea:http://hdl.handle.net/10256/12346
Access Level:acceso embargado
Palabra clave:Ribonuclease A
Ribonucleases
Enzims
Enzymes
Enginyeria de proteïnes
Protein engineering
Descripción
Sumario:Hydrostatic pressure in range 0.1-1.5 GPa is used to modify biological system behaviour mostly in biophysical studies of proteins in solution. Due to specific influence on the system equilibrium high pressure can act as a filter that enables to identify and investigate higher energy protein conformers. The idea of the presented experiments is to examine the behaviour of RNase A molecule under high pressure before and after introduction of destabilizing mutation. For the first time crystal structures of wild-type bovine pancreatic ribonuclease A and its markedly less stable variant modified at position Ile106 were determined at different pressures. X-ray diffraction experiments at high pressure showed that the secondary structure of RNase A is well preserved even beyond 0.67 GPa at room temperature. Detailed structural analysis of ribonuclease A conformation observed under high pressure revealed that pressure influences hydrogen bonds pattern, cavity size and packing of molecule