Correlation of the quaternary structure and enzyme activity in the beta-glucosidase of Spodoptera frugiperda (Sfbgly)

• Grant number: 18/18537-4
• Support type: Scholarships in Brazil - Doctorate (Direct)
• Effective date (Start): December 01, 2018
• Effective date (End): March 31, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal researcher: Sandro Roberto Marana
• Grantee: Felipe Akihiro Melo Otsuka
• Home Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 19/25955-0 - Reconstruction of Ancient Enzyme Sequences to Understand Cancer: Evolvability and Drug Resistance of Abl1 Tyrosine kinase, BE.EP.DD

Abstract

The beta-glycosidases GH1 displays a characteristic tertiary structure, the (beta/alpha)8 barrel, but very few studies evaluate the influence of their quaternary structure on the catalytic activity and function. Thus, the goal of this project is to characterize the quaternary structure of the beta-glycosidase from Spodoptera frugiperda (Sfbgly; O61594; PDB 5CG0) with emphasis on the dimeric interface connecting the formation of this oligomer with effects on the enzyme kinetic parameters. Therefore, mutants (F155N, F155A and a sextuple mutant) were designed targeting the residues within the dimer interface of the Sfbgly. So, it is expected that the mutant enzymes will be monomers and the wild-type dimers. Then, biophysical assays (SEC-MALS, DLS and SAXS) will be used to dissect the oligomer state of these proteins. In addition, enzyme assays will be performed to evaluate the steady-state kinetic parameters of the wild-type and mutants. In brief, these data will allow us to correlate the enzymatic activity with the Sfbgly quaternary structure. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

OTSUKA, FELIPE A. M.; CHAGAS, RAFAEL S.; ALMEIDA, VITOR M.; MARANA, SANDRO R.. Homodimerization of a glycoside hydrolase family GH1 beta-glucosidase suggests distinct activity of enzyme different states. Protein Science, v. 29, n. 9, JUL 2020. (18/18537-4, 18/25952-8, 16/22365-9)