A comparative molecular dynamics study of thermophilic and mesophilic β-fructosidase enzymes

• Published in: Journal of molecular modeling 2015-09, Vol.21 (9), p.228-228, Article 228
• Main Authors: Mazola, Yuliet, Guirola, Osmany, Palomares, Sucel, Chinea, Glay, Menéndez, Carmen, Hernández, Lázaro, Musacchio, Alexis
• Format: Article
• Language: English
• Subjects: Arabidopsis - enzymology; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; beta-Fructofuranosidase - genetics; beta-Fructofuranosidase - metabolism; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Computer Appl. in Life Sciences; Computer Applications in Chemistry; Molecular Dynamics Simulation; Molecular Medicine; Original Paper; Protein Conformation; Sequence Alignment; Temperature; Theoretical and Computational Chemistry; Thermotoga maritima - enzymology
• ISSN: 1610-2940; 0948-5023
• DOI: 10.1007/s00894-015-2772-4

Arabidopsis thaliana cell wall invertase 1 (AtcwINV1) and Thermotoga maritima β-fructosidase (BfrA) are among the best structurally studied members of the glycoside hydrolase family 32. Both enzymes hydrolyze sucrose as the main substrate but differ strongly in their thermal stability. Mesophilic AtcwINV1 and thermophilic BfrA have divergent sequence similarities in the N-terminal five bladed β-propeller catalytic domain (31 %) and the C-terminal β-sandwich domain (15 %) of unknown function. The two enzymes were subjected to 200 ns molecular dynamics simulations at 300 K (27 °C) and 353 K (80 °C). Regular secondary structure regions, but not loops, in AtcwINV1 and BfrA showed no significant fluctuation differences at both temperatures. BfrA was more rigid than AtcwINV1 at 300 K. The simulation at 353 K did not alter the structural stability of BfrA, but did increase the overall flexibility of AtcwINV1 exhibiting the most fluctuating regions in the β-propeller domain. The simulated heat treatment also increased the gyration radius and hydrophobic solvent accessible surface area of the plant enzyme, consistent with the initial steps of an unfolding process. The preservation of the conformational rigidity of BfrA at 353 K is linked to the shorter size of the protein loops. Shortening of BfrA loops appears to be a key mechanism for thermostability.