Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/1065
Type: Artigo de periódico
Title: Molecular Basis of the Thermostability and Thermophilicity of Laminarinases: X-ray Structure of the Hyperthermostable Laminarinase from Rhodothermus marinus and Molecular Dynamics Simulations
Author: BLEICHER, Lucas
PRATES, Erica T.
GOMES, Thiago C. F.
SILVEIRA, Rodrigo L.
NASCIMENTO, Alessandro S.
ROJAS, Adriana L.
GOLUBEV, Alexander
MARTINEZ, Leandro
SKAF, Munir S.
POLIKARPOV, Igor
Abstract: Glycosyl hydrolases are enzymes capable of breaking the glycosidic linkage of polysaccharides and have considerable industrial and biotechnological applications. Driven by the later applications, it is frequently desirable that glycosyl hydrolases display stability and activity under extreme environment conditions, such as high temperatures and extreme pHs. Here, we present X-ray structure of the hyperthermophilic laminarinase from Rhodothermus marinus (RmLamR) determined at 1.95 angstrom resolution and molecular dynamics simulation studies aimed to comprehend the molecular basis, for the thermal stability of this class of enzymes. As most thermostable proteins, RmLamR contains a relatively large number of salt bridges, which are not randomly distributed on the structure. On the contrary, they form clusters interconnecting beta-sheets of the catalytic domain. Not all salt bridges, however, are beneficial for the protein thermostability: the existence of charge-charge interactions permeating the hydrophobic core of the enzymes actually contributes to destabilize the structure by facilitating water penetration into hydrophobic cavities, as can be seen in the case of mesophilic enzymes. Furthermore, we demonstrate that the mobility of the side-chains is perturbed differently in each class of enzymes. The side-chains of loop residues surrounding the catalytic cleft in the mesophilic laminarinase gain mobility and obstruct the active site at high temperature. By contrast, thermophilic laminarinases preserve their active site flexibility, and the active-site cleft remains accessible for recognition of polysaccharide substrates even at high temperatures. The present results provide structural insights into the role played by salt-bridges and active site flexibility on protein thermal stability and may be relevant for other classes of proteins, particularly glycosyl hydrolases.
Country: Estados Unidos
Editor: AMER CHEMICAL SOC
Citation: JOURNAL OF PHYSICAL CHEMISTRY B, v.115, n.24, p.7940-7949, 2011
Rights: fechado
Identifier DOI: 10.1021/jp200330z
Address: http://dx.doi.org/10.1021/jp200330z
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Date Issue: 2011
Appears in Collections:IQ - Artigos e Materiais de Revistas Científicas

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