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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Molecular Basis of the Thermostability and Thermophilicity of Laminarinases: X-ray Structure of the Hyperthermostable Laminarinase from Rhodothermus marinus and Molecular Dynamics Simulations

Texto completo
Autor(es):
Bleicher, Lucas [1] ; Prates, Erica T. [2] ; Gomes, Thiago C. F. [2] ; Silveira, Rodrigo L. [2] ; Nascimento, Alessandro S. [1] ; Rojas, Adriana L. [1, 3] ; Golubev, Alexander [4] ; Martinez, Leandro [1] ; Skaf, Munir S. [2] ; Polikarpov, Igor [1]
Número total de Autores: 10
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Phys Sao Carlos, BR-13560970 Sao Carlos, SP - Brazil
[2] State Univ Campinas UNICAMP, Inst Chem, BR-13084862 Campinas, SP - Brazil
[3] Ctr Cooperat Res Biosci BioGUNE, Struct Biol Unit, Derio 48160 - Spain
[4] Petersburg Nucl Phys Inst, St Petersburg 188300 - Russia
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: Journal of Physical Chemistry B; v. 115, n. 24, p. 7940-7949, JUN 23 2011.
Citações Web of Science: 18
Resumo

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. (AU)

Processo FAPESP: 10/08680-2 - Aspectos moleculares da degradação de biomassa lignocelulósica: dinâmica de enzimas e nanoarquitetura de paredes celulares de plantas
Beneficiário:Rodrigo Leandro Silveira
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 10/18849-4 - Purificação e caracterização de hidrolases de glicosídeos, produzidas pelo fungo filamentoso Trichoderma harzianum, com perspectiva de aplicabilidade dessas enzimas em coquetéis para hidrólise da biomassa e produção de bioetanol
Beneficiário:Igor Polikarpov
Linha de fomento: Auxílio à Pesquisa - Pesquisador Visitante - Internacional
Processo FAPESP: 09/14107-6 - Dinâmica molecular de hidrolases para sacarificação de celulose e proteínas correlatas
Beneficiário:Érica Teixeira Prates
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 10/16947-9 - Estrutura, dinâmica e função em proteínas: simulação computacional e desenvolvimento de algoritmos
Beneficiário:Leandro Martinez
Linha de fomento: Auxílio à Pesquisa - Regular