Probing protein-sugar interactions.
We have investigated the partial specific volumes (2) (ml/g), hydration, and cosolvent interactions of rabbit muscle aldolase by equilibrium sedimentation in the analytical ultracentrifuge and by direct density increment (partial differential/partial differentialc(2))(mu) measurements over a range of sugar concentrations and temperature. In a series of sugars increasing in size, glucose, sucrose, raffinose, and alpha-cyclodextrin, (partial differential/ partial differentialc(2))(mu) decreases linearly with the solvent density rho(0). These sugar cosolvents do not interact with the protein; however, the interaction parameter B(1) (g water/g protein) mildly increases with increasing sugar size. The experimental B(1) values are smaller than values calculated by excluded volume (rolling ball) considerations. B(1) relates to hydration in this and in other instances studied. It decreases with increasing temperature, leading to an increase in (2) due to reduced water of hydration electrostriction. The density increments (partial differential/ partial differentialc(2))(mu), however, decrease in concave up form in the case of glycerol and in concave down form for trehalose, leading to more complex behavior in the case of carbohydrates playing a biological role as osmolytes and antifreeze agents. A critical discussion, based on the thermodynamics of multicomponent solutions, is presented.
Ebel
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Institut de Biologie Structurale, CEA-CNRS, F-38027 Grenoble Cedex 1, France.
Eisenberg
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Ghirlando
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D000818:Animals; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry*; D003505:Cyclodextrins / Q000737:chemistry; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry* / Q000378:metabolism*; D005947:Glucose / Q000737:chemistry; D005990:Glycerol; D007700:Kinetics; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000201:enzymology; D011817:Rabbits; D011887:Raffinose / Q000737:chemistry; D012997:Solvents; D013395:Sucrose / Q000737:chemistry; D013816:Thermodynamics; D014199:Trehalose; D014461:Ultracentrifugation / Q000379:methods; D047391:alpha-Cyclodextrins
D002241:Carbohydrates; D003505:Cyclodextrins; D012997:Solvents; D047391:alpha-Cyclodextrins; D013395:Sucrose; D014199:Trehalose; D005634:Fructose-Bisphosphate Aldolase; D005947:Glucose; D011887:Raffinose; D005990:Glycerol; C032613:alpha-cyclodextrin