Recombinant human pancreatic ribonuclease produced in e. coli: Importance of the amino-terminal sequence

Junichiro Futami, Masaharu Seno, Megumi Kosaka, Hiroko Tada, Satimaru Seno, Hidenori Yamada

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)


Human pancreatic ribonuclease I (hRNase 1) in the mature form has been produced in E.coli using T7 expression system. The recombinant hRNase 1 protein was solubilized from the inclusion bodies, refolded in glutathione redox system, and purified through chromatographic procedures by utilizing cation-exchange and reversed-phase columns. The ribonucleolytic activity of recombinant hRNase 1 was examined on yeast RNA and cytidylyl-3′,5′-adenosine revealing the distinctive ribonucleolytic activity. The activity was perfectly inhibited by human placental RNase inhibitor. Truncation of 7 amino acid residues in the amino-terminal sequence resulted in much reduction in ribonucleolytic activity and in affinity to human placental RNase inhibitor with the disintegration of secondary structures of the protein observed by circular dichroism spectra. The present study has revealed the important contribution of the amino-terminal sequence of hRNase I to the characteristics of the protein.

Original languageEnglish
Pages (from-to)406-413
Number of pages8
JournalBiochemical and Biophysical Research Communications
Issue number1
Publication statusPublished - 1995

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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