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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 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

This work has been supported by Grants BFU2009-06935 and BIO2013-43517 from MINECO (Spain)

© Chemical Physics, 2016, vol. 468, p. 53-62

Elsevier

Author: Kurpiewska, Katarzyna
Dziubek, Kamil
Katrusiak, Andrzej
Font, Josep
Ribó i Panosa, Marc
Vilanova i Brugués, Maria
Lewiński, Krzysztof
Date: 2016
Abstract: 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
This work has been supported by Grants BFU2009-06935 and BIO2013-43517 from MINECO (Spain)
Format: application/pdf
ISSN: 0301-0104
Document access: http://hdl.handle.net/10256/12346
Language: eng
Publisher: Elsevier
Collection: MICINN/PN 2010-2012/BFU2009-06935
MINECO/PE 2014-2016/BIO2013-43517-R
Reproducció digital del document publicat a: http://dx.doi.org/10.1016/j.chemphys.2016.01.010
Articles publicats (D-B)
Is part of: © Chemical Physics, 2016, vol. 468, p. 53-62
Rights: Tots els drets reservats
Subject: Ribonuclease A
Ribonucleases
Enzims
Enzymes
Enginyeria de proteïnes
Protein engineering
Title: Structural investigation of ribonuclease A conformational preferences using high pressure protein crystallography
Type: info:eu-repo/semantics/article
Repository: DUGiDocs

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