Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/34632
Title: Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
Authors: Souza, Tatiana A. C. B.
Okamoto, Débora Noma [UNIFESP]
Ruiz, Diego M.
Oliveira, Lilian Caroline Gonçalves [UNIFESP]
Kondo, Marcia Yuri [UNIFESP]
Tersariol, Ivarne Luis dos Santos [UNIFESP]
Juliano, Luiz [UNIFESP]
De Castro, Rosana E.
Gouvea, Iuri Estrada [UNIFESP]
Murakami, Mario T.
Ctr Nacl Pesquisas Energia & Mat
Universidade Federal de São Paulo (UNIFESP)
Univ Nacl Mar del Plata
Univ Mogi das Crazes
Keywords: Natrialba magadii
Extracellular protease
Halophilism
Structure
Stability
Kinetics
Issue Date: 1-Mar-2012
Publisher: Elsevier B.V.
Citation: Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 94, n. 3, p. 798-805, 2012.
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.
URI: http://repositorio.unifesp.br/handle/11600/34632
ISSN: 0300-9084
Other Identifiers: http://dx.doi.org/10.1016/j.biochi.2011.11.011
Appears in Collections:Em verificação - Geral

Files in This Item:
File Description SizeFormat 
WOS000301332200025.pdf1.21 MBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.