Molecular characterization of a family 5 glycoside hydrolase suggests an induced-fit enzymatic mechanism

Author(s):Show less -

Liberato, Marcelo V. ^[1] ; Silveira, Rodrigo L. ^[2] ; Prates, Erica T. ^[2] ; de Araujo, Evandro A. ^[1] ; Pellegrini, Vanessa O. A. ^[1] ; Camilo, Cesar M. ^[1] ; Kadowaki, Marco A. ^[1] ; Neto, Mario de O. ^[3] ; Popov, Alexander ^[4] ; Skaf, Munir S. ^[2] ; Polikarpov, Igor ^[1]

Affiliation

^[1] Univ Sao Paulo, Sao Carlos Inst Phys, BR-13566590 Sao Paulo - Brazil

^[2] Univ Estadual Campinas, Inst Chem, BR-13084862 Sao Paulo - Brazil

^[3] State Univ Sao Paulo, Inst Biosci, BR-18618970 Sao Paulo - Brazil

^[4] European Synchrotron Radiat Facil, CS40220, Grenoble - France

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 6, APR 1 2016.
• Web of Science Citations: 9

Abstract

Glycoside hydrolases (GHs) play fundamental roles in the decomposition of lignocellulosic biomaterials. Here, we report the full-length structure of a cellulase from Bacillus licheniformis (BlCel5B), a member of the GH5 subfamily 4 that is entirely dependent on its two ancillary modules (Ig-like module and CBM46) for catalytic activity. Using X-ray crystallography, small-angle X-ray scattering and molecular dynamics simulations, we propose that the C-terminal CBM46 caps the distal N-terminal catalytic domain (CD) to establish a fully functional active site via a combination of large-scale multidomain conformational selection and induced-fit mechanisms. The Ig-like module is pivoting the packing and unpacking motions of CBM46 relative to CD in the assembly of the binding subsite. This is the first example of a multidomain GH relying on large amplitude motions of the CBM46 for assembly of the catalytically competent form of the enzyme. (AU)

• FAPESP's process: 10/18773-8 - Molecular cloning, expression, purification and structural characterization of endoglucanase from Trichoderma harzianum aiming the development of enzymatic blends for production of lignocellulosic ethanol
• Grantee: Vanessa de Oliveira Arnoldi Pellegrini
• Support type: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 08/56255-9 - Structure and function of enzymes and auxiliary proteins from Trichoderma, active in cell-wall hydrolysis
• Grantee: Igor Polikarpov
• Support type: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 13/08293-7 - CCES - Center for Computational Engineering and Sciences
• Grantee: Munir Salomao Skaf
• Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 13/15582-5 - Multiscale molecular dynamics of glycoside hydrolases and lignocellulosic substrates
• Grantee: Érica Teixeira Prates
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 09/52840-7 - Center of Biological and Industrial Processes for Biofuels - CeProBIO
• Grantee: Igor Polikarpov
• Support type: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 14/10448-1 - Molecular aspects of plant cell wall architecture
• Grantee: Rodrigo Leandro Silveira
• Support type: Scholarships in Brazil - Post-Doctorate