Intraspecies regulation of ribonucleolytic activity

Biochemistry. 2007 Nov 13;46(45):13131-40. doi: 10.1021/bi701521q. Epub 2007 Oct 23.


The evolutionary rate of proteins involved in obligate protein-protein interactions is slower and the degree of coevolution higher than that for nonobligate protein-protein interactions. The coevolution of the proteins involved in certain nonobligate interactions is, however, essential to cell survival. To gain insight into the coevolution of one such nonobligate protein pair, the cytosolic ribonuclease inhibitor (RI) proteins and secretory pancreatic-type ribonucleases from cow (Bos taurus) and human (Homo sapiens) were produced in Escherichia coli and purified, and their physicochemical properties were analyzed. The two intraspecies complexes were found to be extremely tight (bovine Kd = 0.69 fM; human Kd = 0.34 fM). Human RI binds to its cognate ribonuclease (RNase 1) with 100-fold greater affinity than to the bovine homologue (RNase A). In contrast, bovine RI binds to RNase 1 and RNase A with nearly equal affinity. This broader specificity is consistent with there being more pancreatic-type ribonucleases in cows (20) than humans (13). Human RI (32 cysteine residues) also has 4-fold less resistance to oxidation by hydrogen peroxide than does bovine RI (29 cysteine residues). This decreased oxidative stability of human RI, which is caused largely by Cys74, implies a larger role for human RI as an antioxidant. The conformational and oxidative stabilities of both RIs increase upon complex formation with ribonucleases. Thus, RI has evolved to maintain its inhibition of invading ribonucleases, even when confronted with extreme environmental stress. That role appears to take precedence over its role in mediating oxidative damage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biological Evolution
  • Cattle
  • Enzyme Inhibitors / metabolism*
  • Humans
  • Hydrogen Peroxide
  • Models, Molecular
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Placental Hormones / metabolism*
  • Protein Conformation
  • Protein Interaction Domains and Motifs* / genetics
  • Protein Interaction Mapping
  • Ribonuclease, Pancreatic / genetics
  • Ribonuclease, Pancreatic / metabolism*
  • Sequence Alignment


  • Enzyme Inhibitors
  • Placental Hormones
  • placental ribonuclease inhibitor
  • Hydrogen Peroxide
  • Ribonuclease, Pancreatic