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Interaction of cysteine proteinases with recombinant kininogen domain 2, expressed in Escherichia coli
Author(s) -
Ylinenjärvi Karin,
Prasthofer Thomas W.,
Martin Nancy C.,
Björk Ingemar
Publication year - 1995
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(94)01380-j
Subject(s) - recombinant dna , kininogen , cathepsin b , escherichia coli , cathepsin o , papain , biochemistry , chemistry , cathepsin h , cathepsin l , cathepsin , cathepsin a , cysteine , cysteine proteinase inhibitors , microbiology and biotechnology , cystatin , cathepsin l1 , enzyme , biology , kallikrein , gene , cystatin c , apoptosis , programmed cell death , renal function , caspase
The calpain‐binding domain 2 of the kininogens, the major plasma inhibitors of cysteine proteinases, was expressed in Escherichia coli . Expression of soluble protein was optimal at 15°C and was augmented by growing the bacteria in sorbitol and betaine. The recombinant domain showed high affinity ( K i 0.3–1 nM) for cathepsin L and papain, and a somewhat lower affinity ( K i ∼ 15 nM ) for calpain. The binding to cathepsin H was substantially weaker, and no inhibition of actinidin and cathepsin B was detected. The affinity for cathepsin L was comparable to that reported for the domain isolated from plasma L‐kininogen, whereas the affinities for papain and calpain were about tenfold lower. The latter difference may be due to the recombinant domain being nonglycosylated.