Effects of dimethyl sulfoxide on subunit proteins.
The effects of DMSO are thought to result from the formation of hydrogen bonds with proton-donor groups on biopolymers, which are stronger than those formed with water. Since DMSO contains methyl groups, however, effects on hydrophobic bonding in proteins could be expected at higher DMSO levels. Our studies of the effects of DMSO on model subunit proteins can be interpreted in the above terms. At a concentration of 20% or less, DMSO changed glutamate dehydrogenase into the inactive monomer and the effects were fully reversible with the activator (ADP). Higher DMSO levels resulted in irreversible inactivation. The predominant effect noted on beta-glucuronidase was irreversible inactivation by 20% or more DMSO at 37 degrees C. Purified beta-glucuronidase exhibited an activation in 20% DMSO at high substrate levels; this resulted from an apparent substrate inhibition in the absence of DMSO. DMSO inhibited the clotting of fibrinogen by purified thrombin, but the major effect appeared to be due to competition between thrombin and DMSO for binding sites on fibrinogen. These effects appear to be largely due to interactions between DMSO and hydrophobic bonding in fibrinogen, although DMSO also appears to interfere with the aggregation of fibrin monomers through its effects on hydrophilic groups. These results suggest that reversible alterations in protein structure are the major effect of exposure of subunit proteins to low DMSO levels at low temperatues, while irreversible denaturation of subunit proteins may be an appreciable effect a higher temperatures and higher DMSO concentrations.
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D001777:Blood Coagulation; D002417:Cattle; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005340:Fibrinogen / Q000494:pharmacology; D005966:Glucuronidase / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000032:analysis; D006629:Hirudins / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D011489:Protein Denaturation; D011506:Proteins / Q000378:metabolism*; D013696:Temperature; D013917:Thrombin / Q000494:pharmacology
D006629:Hirudins; D011506:Proteins; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D005969:Glutamate Dehydrogenase; D005966:Glucuronidase; D013917:Thrombin; D004121:Dimethyl Sulfoxide