title: Potent inhibition of ribonuclease A by oligo(vinylsulfonic acid).
abstract: Ribonuclease A (RNase A) can make multiple contacts with an RNA substrate. In particular, the enzymatic active site and adjacent subsites bind sequential phosphoryl groups in the RNA backbone through Coulombic interactions. Here, oligomers of vinylsulfonic acid (OVS) are shown to be potent inhibitors of RNase A that exploit these interactions. Inhibition is competitive with substrate and has Ki = 11 pm in assays at low salt concentration. The effect of salt concentration on inhibition indicates that nearly eight favorable Coulombic interactions occur in the RNase A.OVS complex. The phosphonic acid and sulfuric acid analogs of OVS are also potent inhibitors although slightly less effective. OVS is also shown to be a contaminant of MES and other buffers that contain sulfonylethyl groups. Oligomers greater than nine units in length can be isolated from commercial MES buffer. Inhibition by contaminating OVS is responsible for the apparent decrease in catalytic activity that has been observed in assays of RNase A at low salt concentration. Thus, OVS is both a useful inhibitor of RNase A and a potential bane to chemists and biochemists who use ethanesulfonic acid buffers.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
authors.forename: Bryan D
authors.initials: BD
authors.lastname: Smith
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Soellner
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Raines
mesh_terms: D002021:Buffers; D002384:Catalysis; D005030:Ethylenes / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors* / Q000737:chemistry; D012492:Salts / Q000494:pharmacology; D013451:Sulfonic Acids / Q000494:pharmacology*
chemical_list: D002021:Buffers; D005030:Ethylenes; D012492:Salts; D013451:Sulfonic Acids; D012259:Ribonuclease, Pancreatic; C085726:ethylenesulfonic acid