title: The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 A resolution, and a comparison with related enzymes.
abstract: Cellulose is the most abundant polymer in the biosphere. Although generally resistant to degradation, it may be hydrolysed by cellulolytic organisms that have evolved a variety of structurally distinct enzymes, cellobiohydrolases and endoglucanases, for this purpose. Endoglucanase I (EG I) is the major endoglucanase produced by the cellulolytic fungus Trichoderma reesei, accounting for 5 to 10% of the total amount of cellulases produced by this organism. Together with EG I from Humicola insolens and T. reesei cellobiohydrolase I (CBH I), the enzyme is classified into family 7 of the glycosyl hydrolases, and it catalyses hydrolysis with a net retention of the anomeric configuration. The structure of the catalytic core domain (residues 1 to 371) of EG I from T. reesei has been determined at 3.6 A resolution by the molecular replacement method using the structures of T. reesei CBH I and H. insolens EG I as search models. By employing the 2-fold non-crystallographic symmetry (NCS), the structure was refined successfully, despite the limited resolution. The final model has an R-factor of 0.201 (Rfree 0.258). The structure of EG I reveals an extended, open substrate-binding cleft, rather than a tunnel as found in the homologous cellobiohydrolase CBH I. This confirms the earlier proposal that the tunnel-forming loops in CBH I have been deleted in EG I, which has resulted in an open active site in EG I, enabling it to function as an endoglucanase. Comparison of the structure of EG I with several related enzymes reveals structural similarities, and differences that relate to their biological function in degrading particular substrates. A possible structural explanation of the drastically different pH profiles of T. reesei and H. insolens EG I is proposed.
authors.affiliation: Biomedical Centre, Uppsala University, Uppsala, SE-751 24, Sweden.
authors.forename: G J
authors.initials: GJ
authors.lastname: Kleywegt
authors.forename: J Y
authors.initials: JY
authors.lastname: Zou
authors.forename: C
authors.initials: C
authors.lastname: Divne
authors.forename: G J
authors.initials: GJ
authors.lastname: Davies
authors.forename: I
authors.initials: I
authors.lastname: Sinning
authors.forename: J
authors.initials: J
authors.lastname: Stâhlberg
authors.forename: T
authors.initials: T
authors.lastname: Reinikainen
authors.forename: M
authors.initials: M
authors.lastname: Srisodsuk
authors.forename: T T
authors.initials: TT
authors.lastname: Teeri
authors.forename: T A
authors.initials: TA
authors.lastname: Jones
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology; D001665:Binding Sites; D002475:Cellobiose / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry*; D002482:Cellulose / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase; D003198:Computer Simulation; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D003904:Mitosporic Fungi / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D011487:Protein Conformation; D015202:Protein Engineering; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014242:Trichoderma / Q000201:enzymology*
chemical_list: D010446:Peptide Fragments; D002475:Cellobiose; D002482:Cellulose; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase