Abstract
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 language | English |
|---|---|
| Pages (from-to) | 406-413 |
| Number of pages | 8 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 216 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Nov 2 1995 |
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ASJC Scopus subject areas
- Molecular Biology
- Cell Biology
- Biophysics
- Biochemistry
Cite this
Recombinant Human Pancreatic Ribonuclease Produced in E. coli : Importance of the Amino-Terminal Sequence. / Futami, Junichiro; Seno, Masaharu; Kosaka, Megumi; Tada, Hiroko; Seno, S.; Yamada, H.
In: Biochemical and Biophysical Research Communications, Vol. 216, No. 1, 02.11.1995, p. 406-413.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Recombinant Human Pancreatic Ribonuclease Produced in E. coli
T2 - Importance of the Amino-Terminal Sequence
AU - Futami, Junichiro
AU - Seno, Masaharu
AU - Kosaka, Megumi
AU - Tada, Hiroko
AU - Seno, S.
AU - Yamada, H.
PY - 1995/11/2
Y1 - 1995/11/2
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0028850277&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028850277&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1995.2638
DO - 10.1006/bbrc.1995.2638
M3 - Article
C2 - 7488119
AN - SCOPUS:0028850277
VL - 216
SP - 406
EP - 413
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -