Cloning and characterization of novel ficolins from the solitary ascidian, Halocynthia roretzi.
Ficolins are animal lectins with collagen-like and fibrinogen-like domains. They are involved in the first line of host defense against pathogens. Human ficolin/P35 as well as mannose-binding lectin (MBL) activates the complement lectin pathway in association with MBL-associated serine proteases. To elucidate the origin and evolution of ficolins, we separated approximately 40 kDa (p40) and approximately 50 kDa (p50) N-acetylglucosamine-binding lectins from hemolymph plasma of the solitary ascidian. Binding assays revealed that p40 recognizes N-acetyl groups in association with a pyranose ring and that p50 recognizes N-acetylglucosamine alone. Based on the amino acid sequences of the proteins, we isolated two clones each of p40 and p50 from the ascidian hepatopancreas cDNA and determined the entire coding sequences of these clones. Because all of the clones contained both collagen-like and fibrinogen-like domains, we concluded that these were homologs of the mammalian ficolin family and designated ascidian ficolins (AsFCNs). The fibrinogen-like domain of the AsFCNs shows 45.4-52.4% amino acid sequence identity with the mammalian ficolin family. A phylogenetic tree of the fibrinogen-like sequences shows that all the fibrinogen-like domains may have evolved from a common ancestor that branched off an authentic fibrinogen. These results suggest that AsFCNs play an important role with respect to ascidian hemolymph lectin activity and the correlation of different functions with binding specificity.
Kenjo
A
A
Department of Biochemistry, Fukushima Medical University, School of Medicine, 1-Hikariga-oka, Fukushima 960-1295, Japan.
Takahashi
M
M
Matsushita
M
M
Endo
Y
Y
Nakata
M
M
Mizuochi
T
T
Fujita
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D037102:Lectins; D008969:Molecular Sequence Data; D010802:Phylogeny; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D014561:Urochordata; D000097604:Ficolins
D002352:Carrier Proteins; D017931:DNA Primers; D006023:Glycoproteins; D037102:Lectins; D000097604:Ficolins