Efficient production of native, biologically active human cystatin C by Escherichia coli

Magnus Abrahamson, Henrik Dalboge, Isleifur Olafsson, Sören Carlsen, Anders Grubb

Research output: Contribution to journalArticlepeer-review

Abstract

A cDNA encoding the mature human cysteine proteinase inhibitor cystatin C was fused to the coding sequence for the Escherichia coli outer membrane protein A signal peptide, and the recombinant gene was expressed in E. coli under the control of the λ PR promoter, an optimized Shine-Dalgarno sequence and the λ cI 857 repressor. When induced at 42°C, such cells expressed large amounts of recombinant cystatin C. The recombinant protein was isolated in high yield and characterized. All physicochemical properties investigated, including the positions of disulfide bonds, indicated that the E. coli derived cystatin C was identical to cystatin C isolated from human biological fluids, except that the proline residue in position three was not hydroxylated. The recombinant protein displayed full biological activity against papain, cathepsin B and dipeptidyl peptidase I.
Original languageEnglish
Pages (from-to)14-18
JournalFEBS Letters
Volume236
Issue number1
DOIs
Publication statusPublished - 1988

Subject classification (UKÄ)

  • Biological Sciences

Keywords

  • Cystatin C
  • Cysteine proteinase inhibitor
  • Gene expression
  • Amyloidosis
  • Cerebral hemorrhage

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