NMR studies on interactions of ribonuclease Sa with Guo-3'-P.

Some features of the interaction of guanyloribonuclease Sa from Streptomyces aureofaciens with its competitive inhibitor Guo-3'-P were investigated by 1H and 31P NMR spectroscopy. The pH dependence of chemical shifts of C(2)-H protons of the histidine residue of the enzyme were analysed, in the absence and presence of Guo-3'-P. This analysis showed that only one of the two histidines of ribonuclease Sa is located in the active site of the enzyme. 31P NMR resonances of the nucleotide and of its complex with the enzyme indicated that this histidine interacts with the phosphate group of the substrate. The possible relationship between the observed perturbation of the NMR titration curve of the active site of histidine and a conformational change in the enzyme molecule at a pH of approximately 7.5 is also discussed.