title: Cloning and characterization of a shrimp clip domain serine protease homolog (c-SPH) as a cell adhesion molecule.
abstract: Clip domain serine protease homologs (c-SPHs) are involved in various innate immune functions in arthropods such as antimicrobial activity, cell adhesion, pattern recognition, opsonization, and regulation of the prophenoloxidase system. In the present study, we cloned a c-SPH cDNA from tiger shrimp (Penaeus monodon) hemocytes. It is 1337 bp in length with a coding region of 1068 bp consisting a protein of 355 amino acid residues. The deduced protein includes one clip domain and one catalytically inactive serine protease-like (SP-like) domain. Its molecular weight is estimated to be 38 kDa with an isoelectric point of 7.9. The predicted cutting site of the signal peptide is located between Gly(21) and Gln(22). We aligned 15 single clip domain SPH protein sequences from 12 arthropod species; the identity of these clip domains is low and that of SP-like domains is from 34% to 46%. The conserved regions are located near the amino acid residues which served as substrate interaction sites in catalytically active serine protease. Phylogenetically, the tiger shrimp c-SPH is most similar to a low molecular mass masquerade-like protein of crayfish, but less similar to c-SPHs in Chelicerata and Insecta. Nested reverse transcription polymerase chain reaction (RT-PCR) revealed that c-SPH mRNA is expressed most in tissues with the highest hemocyte abundance. Antimicrobial and opsonization activities of the molecule were not detected. The expression of c-SPH mRNA in hemocytes was up-regulated at the 12-day post beta-glucan immersion. Recombinant c-SPH could significantly enhance hemocyte adhesion. The result suggests that the shrimp c-SPH protein plays a role in innate immunity.
authors.affiliation: Institute of Zoology, National Taiwan University, Taipei, Taiwan.
authors.forename: Chun-Yu
authors.initials: CY
authors.lastname: Lin
authors.forename: Kuang-Yu
authors.initials: KY
authors.lastname: Hu
authors.forename: Shih-Hu
authors.initials: SH
authors.lastname: Ho
authors.forename: Yen-Ling
authors.initials: YL
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D006434:Hemocytes / Q000276:immunology; D015151:Immunoblotting; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D033561:Penaeidae / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
chemical_list: D015815:Cell Adhesion Molecules; D011994:Recombinant Proteins; D012697:Serine Endopeptidases