Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase

Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase

Author Souza, Tatiana A. C. B. Google Scholar
Okamoto, Débora Noma Autor UNIFESP Google Scholar
Ruiz, Diego M. Google Scholar
Oliveira, Lilian Caroline Gonçalves Autor UNIFESP Google Scholar
Kondo, Marcia Yuri Autor UNIFESP Google Scholar
Tersariol, Ivarne Luis dos Santos Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
De Castro, Rosana E. Google Scholar
Gouvea, Iuri Estrada Autor UNIFESP Google Scholar
Murakami, Mario T. Google Scholar
Institution Ctr Nacl Pesquisas Energia & Mat
Universidade Federal de São Paulo (UNIFESP)
Univ Nacl Mar del Plata
Univ Mogi das Crazes
Abstract Nep (Natrialba magadii extracellular protease) is a halolysin-like peptidase secreted by the haloalkaliphilic archaeon N. magadii that exhibits optimal activity and stability in salt-saturated solutions. in this work, the effect of salt on the function and structure of Nep was investigated. in absence of salt. Nep became unfolded and aggregated, leading to the loss of activity. the enzyme did not recover its structural and functional properties even after restoring the ideal conditions for catalysis. At salt concentrations higher than 1 M (NaCl), Nep behaved as monomers in solution and its enzymatic activity displayed a nonlinear concave-up dependence with salt concentration resulting in a 20-fold activation at 4 M NaCl. Although transition from a high to a low-saline environment (3-1 M NaCl) did not affect its secondary structure contents, it diminished the enzyme stability and provoked large structural rearrangements, changing from an elongated shape at 3 M NaCl to a compact conformational state at 1 M NaCl. the thermodynamic analysis of peptide hydrolysis by Nep suggests a significant enzyme reorganization depending on the environmental salinity, which supports in solution SAXS and DLS studies. Moreover, solvent kinetic isotopic effect (SKIE) data indicates the general acid-base mechanism as the rate-limiting step for Nep catalysis, like classical serine-peptidases. All these data correlate the Nep conformational states with the enzymatic behavior providing a further understanding on the stability and structural determinants for the functioning of halolysins under different salinities. (C) 2011 Elsevier Masson SAS. All rights reserved.
Keywords Natrialba magadii
Extracellular protease
Halophilism
Structure
Stability
Kinetics
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Ministerio de Ciencia, Tecnologia e Innovacion Productiva (MINCyT)
Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Argentina
Grant number Ministerio de Ciencia, Tecnologia e Innovacion Productiva (MINCyT): BR09/04
Date 2012-03-01
Published in Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 94, n. 3, p. 798-805, 2012.
ISSN 0300-9084 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 798-805
Origin http://dx.doi.org/10.1016/j.biochi.2011.11.011
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000301332200025
URI http://repositorio.unifesp.br/handle/11600/34632

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