Journal : Fish and Shellfish Immunology , vol. 15 , p. 13–27 , 2003
Publisher : Elsevier
International Standard Numbers
Printed : 1050-4648
Electronic : 1095-9947
Publication type : Academic article
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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, 65 and 115 kDa, respectively. The high molecular weight alpha-chain of the C3 incorporated C-14-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. Immumohistochemical experiments showed that liver hepatocytes and blood contained C3, and in situ hybridisation experiments revealed that liver hepatocytes expressed C3. (C) 2003 Elsevier Science Ltd. All rights reserved.