High-level expression of enzymatically active bovine leukemia virus proteinase in E. coli

Martin Andreánsky, Olga Hrušková-Heidingsfeldová, Juraj Sedláčeka, Jan Konvalinka, Ivo Bláha, Petr Ječmen, Magda Hořejši, Petr Štrop, Milan Fábry

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

An E. coli plasmid expressing efficiently an artificial precursor of bovine leukemia virus (BLV) proteinase under transcriptional control of the phage T7 promoter was constructed. The expression product accumulates in the induced E. coli cells in the form of insoluble cytoplasmic inclusions. Solubilization of the inclusions and a refolding step yield almost pure and completely self-processed proteinase. Purification to homogeneity was achieved by ion-exchange chromatography and reverse-phase HPLC. On a preparative scale, a high yield of enzymatically active proteinase was obtained. An initial study using a series of synthetic peptide substrates shows a distinct substrate specificity of BLV proteinase.

Original languageEnglish (US)
Pages (from-to)129-132
Number of pages4
JournalFEBS letters
Volume287
Issue number1-2
DOIs
StatePublished - Aug 5 1991
Externally publishedYes

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Keywords

  • Bovine leukemia virus
  • E. coli
  • Polyprotein precursor processing
  • Recombinant product accumulation
  • Retroviral proteinase
  • Synthetic peptide substrate

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Andreánsky, M., Hrušková-Heidingsfeldová, O., Sedláčeka, J., Konvalinka, J., Bláha, I., Ječmen, P., Hořejši, M., Štrop, P., & Fábry, M. (1991). High-level expression of enzymatically active bovine leukemia virus proteinase in E. coli. FEBS letters, 287(1-2), 129-132. https://doi.org/10.1016/0014-5793(91)80032-X