Interpretation of the reversible inhibition of adenosine deaminase by small cosolutes in terms of molecular crowding.
Published results on the inhibitory effects of small cosolutes on adenosine deamination by adenosine deaminase [Kurz, L. C., Weitkamp, E., and Frieden, C. (1987) Biochemistry 26, 3027-3032; Dzingeleski, G., and Wolfenden, R. (1993) Biochemistry 32, 9143-9147] have been reexamined. Results for sucrose, dioxane, methanol, and ethanol are shown to be qualitatively consistent with thermodynamic interpretation in terms of molecular crowding effects arising from the occurrence of a minor increase in enzyme volume and/or asymmetry during the kinetic reaction--a conformational transition that could be either preexisting or ligand induced. Direct evidence for the existence of the putative isomeric transition is provided by active enzyme gel chromatography on Sephadex G-100, which demonstrates a negative dependence of enzyme elution volume upon substrate concentration and is therefore consistent with substrate-mediated conformational changes that favor a larger (or more asymmetric) isomeric state of the enzyme. There are thus experimental grounds for adopting the present description of the inhibitory effects of unrelated cosolutes on the kinetics of adenosine deamination by adenosine deaminase in terms of thermodynamic nonideality.
Lonhienne
T G
TG
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Australia.
Winzor
D J
DJ
D000241:Adenosine / Q000378:metabolism; D000243:Adenosine Deaminase / Q000737:chemistry / Q000378:metabolism*; D058892:Adenosine Deaminase Inhibitors; D000818:Animals; D002417:Cattle; D004146:Dioxanes / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D008956:Models, Chemical; D012996:Solutions; D013395:Sucrose / Q000378:metabolism; D013816:Thermodynamics
D058892:Adenosine Deaminase Inhibitors; D004146:Dioxanes; D012996:Solutions; D000431:Ethanol; D013395:Sucrose; D000243:Adenosine Deaminase; C025223:1,4-dioxane; D000241:Adenosine; D000432:Methanol