Crystal structure of the catalytic core domain of the family 6 cellobiohydrolase II, Cel6A, from Humicola insolens, at 1.92 angstrom resolution

A Varrot, S Hastrup, M Schulein, G J Davies

Research output: Contribution to journalArticlepeer-review

Abstract

The three-dimensional structure of the catalytic core of the family 6 cellobiohydrolase II, Cel6A (CBH II), from Humicola insolens has been determined by X-ray crystallography at a resolution of 1.92 A Angstrom. The structure was solved by molecular replacement using the homologous Trichoderma reesei CBH II as a search model. The H. insolens enzyme displays a high degree of structural similarity with its T. reesei equivalent. The structure features both O- (alpha-linked mannose) and N-linked glycosylation and a hexa-co-ordinate Mg2+ ion. The active-site residues are located within the enclosed tunnel that is typical for cellobiohydrolase enzymes and which may permit a processive hydrolysis of the cellulose substrate. The close structural similarity between the two enzymes implies that kinetics and chain-end specificity experiments performed on the H. insolens enzyme are likely to be applicable to the homologous T. reesei enzyme. These cast doubt on the description of cellobiohydrolases as exoenzymes since they demonstrated that Cel6A (CBH II) shows no requirement for non-reducing chain-ends, as had been presumed. There is no crystallographic evidence in the present structure to support a mechanism involving loop opening, yet preliminary modelling experiments suggest that the active-site tunnel of Ce16A (CBH II) is too narrow to permit entry of a fluorescenylderivatize substrate, known to be a viable substrate for this enzyme.

Original languageEnglish
Pages (from-to)297-304
Number of pages8
JournalBiochemical journal
Volume337
Publication statusPublished - 15 Jan 1999

Keywords

  • SEQUENCE-BASED CLASSIFICATION
  • TRICHODERMA-REESEI
  • GLYCOSYL HYDROLASES
  • MACROMOLECULAR STRUCTURES
  • 3-DIMENSIONAL STRUCTURE
  • ACTIVE-SITE
  • CELLULASES
  • CELLULOSE
  • DEGRADATION
  • SUBSTRATE

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