| Full text | |
| Author(s): |
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]
Total Authors: 10
|
| Affiliation: | [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
Total Affiliations: 4
|
| Document type: | Journal article |
| Source: | Journal of Physical Chemistry B; v. 115, n. 24, p. 7940-7949, JUN 23 2011. |
| Web of Science Citations: | 18 |
| 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. (AU) | |
| FAPESP's process: | 10/08680-2 - Molecular aspectos of lignocellulosic biomass degradation: dynamics of enzymes and plant cell wall nanoarchitecture |
| Grantee: | Rodrigo Leandro Silveira |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 09/14107-6 - Molecular dynamics simulations of hydrolases for saccharification of cellulose and related proteins |
| Grantee: | Érica Teixeira Prates |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 10/18849-4 - Purification and characterization of glycoside hydrolases produced by the filamentous fungus Trichoderma harzianum, with perspective of applicability of these enzymes in cocktails for biomass hydrolysis and bioethanol production |
| Grantee: | Igor Polikarpov |
| Support Opportunities: | Research Grants - Visiting Researcher Grant - International |
| FAPESP's process: | 10/16947-9 - Correlations between dynamics, structure and function in protein: computer simulations and algorithms |
| Grantee: | Leandro Martinez |
| Support Opportunities: | Regular Research Grants |