Unfolding of cardosin A in organic solvents and detection of intermediaries

被引：9

作者：

Sarmento, Ana Cristina ^{[1
]}

Oliveira, Claudia S. ^{[1
]}

Pereira, Anabela ^{[1
]}

Esteves, Valdemar I. ^{[2
]}

Moir, Arthur J. G. ^{[3
]}

Saraiva, Jorge ^{[4
]}

Pires, Euclides ^{[5
,6
]}

Barros, Marlene ^{[5
,7
]}

机构：

[1] Univ Aveiro, Dept Biol, CESAM, P-3810193 Aveiro, Portugal

[2] Univ Aveiro, Dept Quim, CESAM, P-3810193 Aveiro, Portugal

[3] Univ Sheffield, Krebs Inst, Sheffield, S Yorkshire, England

[4] Univ Aveiro, Dept Quim, QOPNA, P-3810193 Aveiro, Portugal

[5] Univ Coimbra, Ctr Neurosci, P-3500 Coimbra, Portugal

[6] Univ Coimbra, Dept Bioquim, Fac Ciencias & Tecnol, P-3500 Coimbra, Portugal

[7] Univ Catolica Portuguesa, Ctr Reg Beiras, P-3504505 Viseu, Portugal

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2009年 / 57卷 / 1-4期

关键词：

Aspartic proteinases; Organic solvents; Folding; Intermediaries; CYNARA-CARDUNCULUS L; ASPARTIC PROTEINASES; LIMITED PROTEOLYSIS; STABILITY; BEHAVIOR; SYSTEMS; INACTIVATION; HYDROLYSIS; KINETICS; FLOWERS;

D O I：

10.1016/j.molcatb.2008.08.004

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 that were 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. (c) 2008 Elsevier B.V. All rights reserved

引用

页码：115 / 122

页数：8