TAILIEUCHUNG - Báo cáo Y học: Purification and characterization of novel kininogens from spotted wolffish and Atlantic cod

Kininogens are multifunctional proteins found so far mainly in mammals. They carry vasoactive kinins as well as participate in defense, blood coagulation and the acute phase response. In this study, novel kininogens were isolated from Atlantic cod (Gadus morhua L.) and spotted wolffish (Anarhichas minor) by papain-affinity chromatography. The molecular mass of cod kininogen determined by MALDITOF mass spectrometry to be kDa and it had pI values of , and . | Eur. J. Biochem. 269 2639-2646 2002 FEBS 2002 doi Purification and characterization of novel kininogens from spotted wolffish and Atlantic cod Anne Ylonen1 Jari Helin1 Jarl Bogwald2 Anu Jaakola1 Ari Rinne3 and Nisse Kalkkinen1 Institute of Biotechnology Protein Chemistry Laboratory University of Helsinki Finland 2Norwegian College of Fishery Science University of Troms0 Norway 3Institute of Medical Biology University of Troms0 Norway Kininogens are multifunctional proteins found so far mainly in mammals. They carry vasoactive kinins as well as participate in defense blood coagulation and the acute phase response. In this study novel kininogens were isolated from Atlantic cod Gadus morhua L. and spotted wolffish Anarhichas minor by papain-affinity chromatography. The molecular mass of cod kininogen determined by MALDI-TOF mass spectrometry to be kDa and it had pI values of and . The molecular mass of wolffish kininogen was kDa and it had pI values of and . Partial amino-acid sequences determined from both kinin- ogens showed clear homology with previously determined kininogen sequences. Both kininogens were found to inhibit cysteine proteinases like papain and ficin but they had no effect on trypsin a serine proteinase. Wolffish kininogen carried a2 3-sialylated biantennary and triantennary N-gly-cans with extensive sialic acid O-acetylation. Cod kininogen carried similar glycan structures but about 1 3 of its glycans carried sulfate at their N-acetylglucosamine units. Keywords Atlantic cod spotted wolffish kininogen N-gly-cosylation O-acetylation. Kininogens are large molecular mass 50-114 kDa cysteine proteinase inhibitors belonging to class 3 of the cystatin superfamily. They are single-chain proteins composed of an N-terminal heavy chain the bradykinin moiety and a C-terminal light chain. The heavy chain and light chain are interlinked by disulfide bridges. Kininogens are further classified .

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