Premium
Efficient production of native, biologically active human cystatin C by Escherichia coli
Author(s) -
Abrahamson Magnus,
Dalbøge Henrik,
Olafsson Isleifur,
Carlsen Søren,
Grubb Anders
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80276-x
Subject(s) - escherichia coli , recombinant dna , cystatin , biochemistry , signal peptide , papain , complementary dna , cysteine , chemistry , microbiology and biotechnology , cathepsin b , biology , peptide , cystatin c , gene , enzyme , renal function
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 P R 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.