Abstract
The structure of the catalytic core of the endoglucanase V (EGV) from Humicola insolens has been determined by the method of multiple isomorphous replacement at 1.5 Angstrom resolution. The final model, refined with X-PLOR and PROLSQ, has a crystallographic R factor of 0.163 (R(free) = 0.240) with deviations from stereochemical target values of 0.012 Angstrom and 0.037 degrees for bonds and angles, respectively. The model was further refined with SHELXL, including anisotropic modelling of the protein-atom temperature factors, to give a final model with an R factor of 0.105 and an R(free) of 0.154. The initial isomorphous replacement electron-density map was poor and uninterpretable but was improved by the use of synchrotron data collected at a wavelength chosen so as to optimize the f '' contribution of the anomalous scattering from the heavy atoms. The structure of H. insolens EGV consists of a six-stranded beta-barrel domain, similar to that found in a family of plant defence proteins, linked by a number of disulfide-bonded loop regions. A long open groove runs across the surface of the enzyme either side of which lie the catalytic aspartate residues. The 9 Angstrom separation of the catalytic carboxylate groups is consistent with the observation that EGV catalyzes the hydrolysis of the cellulose beta(1-->4) links with inversion of configuration at the anomeric C1 atom. This structure is the first representative from the glycosyl hydrolase family 45.
Original language | English |
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Pages (from-to) | 717 |
Number of pages | 11 |
Journal | Acta Crystallographica. Section D, Biological Crystallography |
Volume | 52 |
Publication status | Published - 1 Jan 1996 |
Keywords
- ACID-SEQUENCE SIMILARITIES
- NUCLEAR-MAGNETIC-RESONANCE
- CELLULOSE-BINDING DOMAIN
- ELECTRON-DENSITY MAPS
- C-TERMINAL DOMAIN
- 3-DIMENSIONAL STRUCTURE
- BARLEY SEED
- TRICHODERMA-REESEI
- PROTEIN MODELS
- FAMILIES