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Small-angle X-ray scattering and structural modeling of full-length: cellobiohydrolase I from Trichoderma harzianum

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Abstract

Cellobiohydrolases hydrolyze cellulose releasing cellobiose units. They are very important for a number of biotechnological applications, such as, for example, production of cellulosic ethanol and cotton fiber processing. The Trichoderma cellobiohydrolase I (CBH1 or Cel7A) is an industrially important exocellulase. It exhibits a typical two domain architecture, with a small C-terminal cellulose-binding domain and a large N-terminal catalytic core domain, connected by an O-glycosylated linker peptide. The mechanism by which the linker mediates the concerted action of the two domains remains a conundrum. Here, we probe the protein shape and domain organization of the CBH1 of Trichoderma harzianum (ThCel7A) by small angle X-ray scattering (SAXS) and structural modeling. Our SAXS data shows that ThCel7A linker is partially-extended in solution. Structural modeling suggests that this linker conformation is stabilized by inter- and intra-molecular interactions involving the linker peptide and its O-glycosylations.

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Acknowledgments

This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) via Thematic Process 08/56255-9, grants 2008/05637-9, 2007/08706-9, 2010/16947-9, CeProBio Project (FAPESP 2009/52840-7 and CNPq 490022/2009-0) and INCT Bioetanol (FAPESP/CNPq); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). We also thank the staff of the National Synchrotron Light Laboratory (LNLS, Brazil) for access to the SAXS beamline and others facilities.

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  1. Mario de Oliveira Neto

    Present address: Departamento de Física e Biofísica, Instituto de Biociências de Botucatu, Unesp, Univ Estadual Paulista, Distrito de Rubião Jr. s/n, Botucatu, SP, Brazil

  2. Geraldo Rodrigues Sartori

    Present address: Institute of Chemistry of São Carlos, University of São Paulo, São Carlos, SP, Brazil

  3. Leandro Martínez

    Present address: Institute of Chemistry, State University of Campinas (UNICAMP), Campinas, SP, Brazil

Authors and Affiliations

  1. Grupo de Biotecnologia Molecular, Instituto de Física de São Carlos (IFSC), Universidade de São Paulo (USP), Av. Trabalhador São-carlense, 400, Centro, São Carlos, SP, 13566-570, Brazil

    Leonardo H. F. Lima, Viviane I. Serpa, Flávio R. Rosseto, Geraldo Rodrigues Sartori, Mario de Oliveira Neto, Leandro Martínez & Igor Polikarpov

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  1. Leonardo H. F. Lima
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Lima, L.H.F., Serpa, V.I., Rosseto, F.R. et al. Small-angle X-ray scattering and structural modeling of full-length: cellobiohydrolase I from Trichoderma harzianum . Cellulose 20, 1573–1585 (2013). https://doi.org/10.1007/s10570-013-9933-3

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