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 MALDI-TOF mass spectrometry to be 51.0 kDa and it had pI values of 3.6, 3.9 and 4.4. The molecular mass of wolffish kininogen was 45.8 kDa and it had pI values of 4.1, 4.3, 4.35 and 4.4. Partial amino-acid sequences determined from both kininogens 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 alpha2,3-sialylated biantennary and triantennary N-glycans 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.
Ylönen
Anne
A
Institute of Biotechnology, Protein Chemistry Laboratory, University of Helsinki, Finland. anne.ylonen@helsinki.fi
Helin
Jari
J
Bøgwald
Jarl
J
Jaakola
Anu
A
Rinne
Ari
A
Kalkkinen
Nisse
N
D000595:Amino Acid Sequence; D000818:Animals; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005399:Fishes / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D007704:Kininogens / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D015853:Cysteine Proteinase Inhibitors; D007704:Kininogens