Binding of the cellulose-binding domain of exoglucanase Cex from Cellulomonas fimi to insoluble microcrystalline cellulose is entropically driven.
Isothermal titration microcalorimetry is combined with solution-depletion isotherm data to analyze the thermodynamics of binding of the cellulose-binding domain (CBD) from the beta-1,4-(exo)glucanase Cex of Cellulomonas fimi to insoluble bacterial microcrystalline cellulose. Analysis of isothermal titration microcalorimetry data against two putative binding models indicates that the bacterial microcrystalline cellulose surface presents two independent classes of binding sites, with the predominant high-affinity site being characterized by a Langmuir-type Ka of 6.3 (+/-1.4) x 10(7) M-1 and the low-affinity site by a Ka of 1.1 (+/-0.6) x 10(6) M-1. CBDCex binding to either site is exothermic, but is mainly driven by a large positive change in entropy. This differs from protein binding to soluble carbohydrates, which is usually driven by a relatively large exothermic standard enthalpy change for binding. Differential heat capacity changes are large and negative, indicating that sorbent and protein dehydration effects make a dominant contribution to the driving force for binding.
Creagh
A L
AL
Department of Chemical Engineering, University of British Columbia, Vancouver, Canada.
Ong
E
E
Jervis
E
E
Kilburn
D G
DG
Haynes
C A
CA
D001665:Binding Sites; D002151:Calorimetry; D002482:Cellulose / Q000378:metabolism*; D002846:Chromatography, Affinity; D043364:Endo-1,4-beta Xylanases; D019277:Entropy; D055672:Static Electricity; D014995:Xylosidases / Q000378:metabolism*; D001617:beta-Glucosidase / Q000378:metabolism*
D002482:Cellulose; D014995:Xylosidases; D001617:beta-Glucosidase; D043364:Endo-1,4-beta Xylanases; C090001:XynB xylanase