A fibrinogen-related protein from bay scallop Argopecten irradians involved in innate immunity as pattern recognition receptor.
The family of fibrinogen-related proteins (FREPs) is a group of proteins with fibrinogen-like domains. Many members of this family play important roles as pattern recognition receptors in innate immune responses. The cDNA of bay scallop Argopecten irradians FREP (designated as AiFREP) was cloned by rapid amplification of cDNA ends (RACE) method based on the expressed sequence tag (EST). The full-length cDNA of AiFREP was of 990bp. The open reading frame encoded a polypeptide of 251 amino acids, including a signal sequence and a 213 amino acids fibrinogen-like domain. The fibrinogen-like domain of AiFREP was highly similar to those of mammalian ficolins and other FREPs. The temporal expression of AiFREP mRNA in hemolymph was examined by fluorescent quantitative real-time PCR. The mRNA level of scallops challenged by Listonella anguillarum was significantly up-regulated, peaked to 9.39-fold at 9h after stimulation, then dropped back to 4.37-fold at 12h, while there was no significant change in the Micrococcus luteus challenged group in all periods of treatment. The function of AiFREP was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli Rosetta gami (DE3). The recombinant AiFREP (rAiFREP) agglutinated chicken erythrocytes and human A, B, O-type erythrocytes. The agglutinating activities were calcium-dependent and could be inhibited by acetyl group-containing carbohydrates. rAiFREP also agglutinated Gram-negative bacteria E. coli JM109, L. anguillarum and Gram-positive bacteria M. luteus in the presence of calcium ions. These results collectively suggested that AiFREP functions as a pattern recognition receptor in the immune response of bay scallop and contributed to nonself recognition in invertebrates, which would also provide clues for elucidating the evolution of the lectin pathway of the complement system.
Zhang
Huan
H
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, Shandong, China.
Wang
Lingling
L
Song
Linsheng
L
Song
Xiaoyan
X
Wang
Bo
B
Mu
Changkao
C
Zhang
Ying
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D005340:Fibrinogen / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006386:Hemagglutination Tests; D007113:Immunity, Innate / Q000276:immunology*; D008024:Ligands; D008969:Molecular Sequence Data; D049897:Pectinidae / Q000235:genetics* / Q000276:immunology*; D011485:Protein Binding; D051192:Receptors, Pattern Recognition / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors
D008024:Ligands; D051192:Receptors, Pattern Recognition; D011994:Recombinant Proteins; D005340:Fibrinogen