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

Abrahamson, Magnus LU ; Dalboge, Henrik; Olafsson, Isleifur; Carlsen, Sören and Grubb, Anders LU (1988) In FEBS Letters 236(1). p.14-18
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cystatin C, Cysteine proteinase inhibitor, Gene expression, Amyloidosis, Cerebral hemorrhage
in
FEBS Letters
volume
236
issue
1
pages
14 - 18
publisher
Wiley-Blackwell
external identifiers
  • scopus:0023684074
ISSN
1873-3468
DOI
10.1016/0014-5793(88)80276-X
language
English
LU publication?
yes
id
de2dcaf2-e922-410c-8866-4b55c5f3be0a (old id 1104385)
date added to LUP
2008-08-08 09:44:02
date last changed
2017-08-06 04:29:16
@article{de2dcaf2-e922-410c-8866-4b55c5f3be0a,
  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.},
  author       = {Abrahamson, Magnus and Dalboge, Henrik and Olafsson, Isleifur and Carlsen, Sören and Grubb, Anders},
  issn         = {1873-3468},
  keyword      = {Cystatin C,Cysteine proteinase inhibitor,Gene expression,Amyloidosis,Cerebral hemorrhage},
  language     = {eng},
  number       = {1},
  pages        = {14--18},
  publisher    = {Wiley-Blackwell},
  series       = {FEBS Letters},
  title        = {Efficient production of native, biologically active human cystatin C by Escherichia coli},
  url          = {http://dx.doi.org/10.1016/0014-5793(88)80276-X},
  volume       = {236},
  year         = {1988},
}