title: The spotted wolffish (Anarhichas minor Olafsen) complement component C3: isolation, characterisation and tissue distribution.
abstract: The complement component C3 was isolated from spotted wolffish (Anarhichas minor Olafsen) serum by polyethylene glycol precipitation, anion exchange chromatography and gel filtration. Silver staining in SDS-PAGE and rabbit anti-wolffish C3 antiserum used in Western blotting revealed that spotted wolffish C3 contains two polypeptide chains, M(r)65 and 115kDa, respectively. The high molecular weight alpha-chain of the C3 incorporated 14C-methylamine suggesting that it contained a reactive thioester group. The deduced amino acid sequence, after screening a liver cDNA expression library, showed that the wolffish C3 contained key amino acids for binding C3 convertase, factor H, I and properdin. Also, high degree of homology to other vertebrate C3 was found in the beta-alpha junction site. Phylogenetic tree analysis indicated that the Japanese flounder and spotted wolffish that belong to order pleuronectiformes and perciformes, respectively, are phylogenetically close species. Immunohistochemical experiments showed that liver hepatocytes and blood contained C3, and in situ hybridisation experiments revealed that liver hepatocytes expressed C3.
authors.affiliation: Department of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, N9037 Tromsø, Norway.
authors.forename: Torill K
authors.initials: TK
authors.lastname: Abelseth
authors.forename: Klara
authors.initials: K
authors.lastname: Stensvåg
authors.forename: Sigrun
authors.initials: S
authors.lastname: Espelid
authors.forename: Randi
authors.initials: R
authors.lastname: Nygaard
authors.forename: Terje
authors.initials: T
authors.lastname: Ellingsen
authors.forename: Jarl
authors.initials: J
authors.lastname: Bøgwald
authors.forename: Roy A
authors.initials: RA
authors.lastname: Dalmo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western / Q000662:veterinary; D002846:Chromatography, Affinity / Q000662:veterinary; D003176:Complement C3 / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry / Q000662:veterinary; D017403:In Situ Hybridization / Q000662:veterinary; D007525:Isoelectric Focusing / Q000662:veterinary; D008969:Molecular Sequence Data; D010473:Perciformes / Q000145:classification / Q000235:genetics / Q000276:immunology*; D010802:Phylogeny; D011817:Rabbits
chemical_list: D003176:Complement C3; D018076:DNA, Complementary; D007106:Immune Sera