title: Coelomocytes express SpBf, a homologue of factor B, the second component in the sea urchin complement system.
abstract: A homologue of factor B, SpBf, has been cloned and sequenced from an LPS-activated coelomocyte cDNA library from the purple sea urchin, Strongylocentrotus purpuratus. The deduced amino acid sequence and domain structure show significant similarity to the vertebrate Bf/C2 family proteins. SpBf is a mosaic protein, composed of five short consensus repeats, a von Willebrand Factor domain, and a serine protease domain. It has a deduced molecular mass of 91 kDa, with a conserved cleavage site for a putative factor D protease. It has ten consensus recognition sites for N-linked glycosylation. Amino acids involved in both Mg2+ binding and in serine protease activity in the vertebrate C2/Bf proteins are conserved in SpBf. Phylogenetic analysis of SpBf indicates that it is the most ancient member of the vertebrate Bf/C2 family. Additional phylogenetic analysis of the SCRs indicates that five SCRs in SpBf may be ancestral to three SCRs, which is the typical pattern in the vertebrate Bf/C2 proteins. RNA gel blots show that SpBf transcripts are 5.5 kb and are specifically expressed in coelomocytes. Genome blots suggest that the SpBf gene (Sp152) is single copy gene per haploid genome. This is the second complement component to be identified from the sea urchin, and, with the sea urchin C3 homologue, these two components may be part of a simple complement system that is homologous to the alternative pathway in higher vertebrates.
authors.affiliation: Department of Biological Sciences, Institute of Biomedical Sciences, George Washington University, Washington, DC 20052, USA. csmith@gwu.edu
authors.forename: L C
authors.initials: LC
authors.lastname: Smith
authors.forename: C S
authors.initials: CS
authors.lastname: Shih
authors.forename: S G
authors.initials: SG
authors.lastname: Dachenhausen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003170:Complement Pathway, Alternative; D003165:Complement System Proteins; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005796:Genes; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D007107:Immune System / Q000166:cytology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012617:Sea Urchins / Q000166:cytology / Q000276:immunology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014714:Vertebrates / Q000235:genetics / Q000276:immunology
chemical_list: D018076:DNA, Complementary; D006023:Glycoproteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; C116571:SpBf protein, Strongylocentrotus purpuratus; D003165:Complement System Proteins; D004247:DNA