Cosolvent-buffer mixtures as models for the cytoplasmic mileu: the enzymology of adenosine aminohydrolase.
Adenosine aminohydrolase from calf intestinal mucosa is sensitive to changes in its environment produced by small mole fractions of dimethylsulfoxide (DMSO). At a mole fraction of 0.1 where the dielectric constant is lowered from that of 78 of neat water to about 76.5, Vmax was reduced by 65% and affinity for substrate (adenosine) and the two competitive inhibitors, insine and N6-benzyladenosine, was decreased markedly. However, this decreased affinity was such that Ki/Km remained virtually constant for both inhibitors. DMSO itself showed the kinetics of a mixed inhibitor with Ki decreasing with increasing mole fraction. This cosolvent also decreased the heat stability of the enzyme which suggests that enzyme conformation is altered by DMSO. Comparison of data in the presence of DMSO with previously obtained data with dioxane shows that heat stability as well as Vmax, at a given value of dielectric constant, is independent of the amount or nature of cosolvent used to achieve that dielectric constant. However, cosolvent induced changes in Ki indicate that colligative as well as dielectric constant effects contribute to the observed changes in kinetic behavior. These experiments may be considered as models for the behavior of enzymes in the medium of lowered dielectric constant expected in the vicinity of cytoplasmic membranes. The results indicate that in such an environment, adenosine aminohydrolase would be expected to be less efficient a catalyst, but equally susceptible to product inhibition, as compared to media of dielectric constant approaching that of water.
D000243:Adenosine Deaminase / Q000378:metabolism*; D000818:Animals; D002021:Buffers; D002417:Cattle; D003593:Cytoplasm / Q000201:enzymology; D004121:Dimethyl Sulfoxide; D004355:Drug Stability; D004553:Electric Conductivity; D006358:Hot Temperature; D007413:Intestinal Mucosa / Q000201:enzymology; D007700:Kinetics; D008954:Models, Biological; D009700:Nucleoside Deaminases / Q000378:metabolism*; D012997:Solvents; D013816:Thermodynamics
D002021:Buffers; D012997:Solvents; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase; D004121:Dimethyl Sulfoxide