Abstract
Limited proteolysis (papain) of the cellobiohydrolase I (CBH I, 65 kDa) from Trichoderma reesei led to the seperation of two functional domains: a core protein (55 kDa) containing the active site, and a C-terminal glycopeptide (10 kDa) implicated in binding to the insoluble matrix (cellulose). The quaternary structures of the intact CBH I and its core in solution are now compared by small angle X-ray scattering (SAXS) measurements. The molecular parameters derived for the core (Rg=2.09 nm, Dmax=6.5 nm) and for the intact enzyme (Rg=4.27 nm, Dmax=18 nm) indicate very different shapes. The resulting models show a “tadpole”-like structure for the intact enzyme where the isotropic part coincides with the core protein and the flexible tail part should be identified with the C-terminal glycopeptide. Thus in this enzyme, functional differentiation is reflected in structural peculiarities.
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Abbreviations
- SAXS:
-
small angle X-ray scattering
- SDS-PAGE:
-
SDS-polyacrylamide gel electrophoresis
- IEF-PAG:
-
polyacrylamide gel isoelectric focusing; cellobiohydrolase (CBH, 1,4-β-glucan cellobio hydrolase (E.C.3.2.1.91))
- Dmax :
-
maximum diameter
- Rg:
-
radius of gyration
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Abuja, P.M., Schmuck, M., Pilz, I. et al. Structural and functional domains of cellobiohydrolase I from trichoderma reesei . Eur Biophys J 15, 339–342 (1988). https://doi.org/10.1007/BF00254721
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DOI: https://doi.org/10.1007/BF00254721