Please use this identifier to cite or link to this item:
http://hdl.handle.net/10773/6785
Title: | Unfolding of cardosin A in organic solvents and detection of intermediaries |
Author: | Sarmento, Ana Cristina Oliveira, Cláudia S. Pereira, Anabela Esteves, Valdemar I. Moir, Arthur J.G. Moir Saraiva, Jorge Pires, Euclides Barros, Marlene |
Keywords: | Aspartic proteinases Organic solvents Folding Intermediaries |
Issue Date: | 2009 |
Publisher: | Elsevier |
Abstract: | In the present study the relationship between conformational stability and enzymatic activity in the presence of organic solvents (OS) was investigated. We have found that cardosin A, the model protease investigated in this work, inactivates through a biphasic mechanism, which is incipient in aqueous buffer and becomes visible in the presence of hydrophilic OS. In fact, in OS this inactivation originates stable intermediaries thatwere detected in acetonitrile. This biphasic mechanism can be described in two phases: an initial one, where OS induce alterations that affect the active site cleft and global mobility, but with little interference on the global protein conformation, and, a second phase, where there is a global change in protein conformation with concomitant activity loss. It is shown that in the presence of hydrophilic OS there is a larger mobility of the enzyme, revealed by limited proteolysis, probably due to a weakening of hydrophobic interactions within the protein core. |
Peer review: | yes |
URI: | http://hdl.handle.net/10773/6785 |
DOI: | 10.1016/j.molcatb.2008.08.004 |
ISSN: | 1381-1177 |
Appears in Collections: | DQ - Artigos QOPNA - Artigos |
Files in This Item:
File | Description | Size | Format | |
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Ana Cristina.pdf | 627.28 kB | Adobe PDF |
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