Efficient production of native, biologically active human cystatin C by Escherichia coli
(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:
https://lup.lub.lu.se/record/1104385
- author
- Abrahamson, Magnus LU ; Dalboge, Henrik ; Olafsson, Isleifur ; Carlsen, Sören and Grubb, Anders LU
- organization
- publishing date
- 1988
- 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
- 2016-04-01 16:01:20
- date last changed
- 2021-06-13 03:10:16
@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}}, keywords = {{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}}, doi = {{10.1016/0014-5793(88)80276-X}}, volume = {{236}}, year = {{1988}}, }