Skip to main content
Log in

A novel serine protease with clip domain from scallop Chlamys farreri

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The serine proteases with clip domain are involved in various innate immune functions in invertebrate such as antimicrobial activity, cell adhesion, pattern recognition and regulation of the prophenoloxidase system. A serine protease with clip-domain cDNA (Cf SP) was obtained by Expressed sequence taggings (ESTs) method and rapid amplification of cDNA ends (RACE). The Cf SP full-length cDNA was of 1,152 bp, including a 5′-terminal untranslated region (UTR) of 63 bp, a 3′-terminal UTR of 81 bp with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, and an open reading frame of 1,008 bp encoding a polypeptide of 336 amino acids with a putative signal peptide of 19 amino acids. The deduced amino acid sequence of Cf SP contained an amino-terminal clip domain with three disulfide bonds formed six conserved Cys residues, a carboxyl-terminal trypsin-like domain with the conserved His–Asp–Ser catalytic triad, and a low complexity linker sequence. The Cf SP was strongly expressed in hemocytes and the mRNA expression of Cf SP was up-regulated and increased 3.2-fold and 2.6-fold at 16 h after injection of Vibrio anguillarum and Micrococcus luteus. The results suggested that Cf SP gene might be involved in immune response of Gram-negative and Gram-positive microbial infection in scallop

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Guo X, Ford SE, Zhang F (1999) Molluscan aquaculture in China. J Shellfish Res 18:19–31

    Google Scholar 

  2. Wang Y, Xiang J (1999) Studies on causation of the mass mortality of Chlamys farreri. Oceanol Limnol Sinica 30:770–774, (in Chinese with English abstract)

    Google Scholar 

  3. He G, Li Y, Song W, Wang C, Huang J, Wang X (2003) The relationship between pathogenic infection status and mortality of the scallop Chlamys farreri. J Fish China 27:273–277

    Google Scholar 

  4. Hoffmann JA, Reichhart JM (2002) Drosophila innate immunity, an evolutionary perspective. Nat Immunol 3:121–126

    Article  PubMed  CAS  Google Scholar 

  5. Hoffmann JA (2003) The immune response of Drosophila. Nature 426:33–38

    Article  PubMed  CAS  Google Scholar 

  6. Seo J, Crawford JM, Stone KL, Noga EJ (2005) Purification of a novel arthropod defensin from the American oyster, Crassostrea virginica. Biochem Biophys Res Commun 338:1998–2004

    Article  PubMed  CAS  Google Scholar 

  7. Zhao J, Song L, Li C, Zou H, Ni D, Wang W, Xu W (2007) Molecular cloning of an invertebrate goose-type lysozyme gene from Chlamys farreri, and lytic activity of the recombinant protein. Mol Immunol 44:1198–1208

    Article  PubMed  CAS  Google Scholar 

  8. Medzhitov R, Janeway CA Jr (1997) Innate immunity, impact on the adaptive immune response. Curr Opin Immunol 9:4–9

    Article  PubMed  CAS  Google Scholar 

  9. Medzhitov R, Janeway CA Jr (2002) Decoding the patterns of self and nonself by the innate immune system. Science 296:298–300

    Article  PubMed  CAS  Google Scholar 

  10. Michel T, Reichhart JM, Hoffmann JA, Royet J (2001) Drosophila Toll is activated by Gram-positive bacteria through a circulating peptidoglycan recognition protein. Nature 414:756–759

    Article  PubMed  CAS  Google Scholar 

  11. Christophides GK, Zdobnov E, Barillas-Mury C, Birney E, Blandin S, Blass C, Brey PT, Collins FH, Danielli A, Dimopoulos G, Hetru C, Hoa NT, Hoffmann JA, Kanzok SM, Letunic I, Levashina EA, Loukeris TG, Lycett G, Meister S, Michel K, Moita LF, Muller HM, Osta MA, Paskewitz SM, Reichhart JM, Rzhetsky A, Troxler L, Vernick KD, Vlachou D, Volz J, von Mering C, Xu J, Zheng L, Bork P, Kafatos FC (2002) Immunity-related genes and gene families in Anopheles gambiae. Science 298:159–165

    Article  PubMed  CAS  Google Scholar 

  12. Dimopoulos G, Muller HM, Levashina EA, Kafatos FC (2001) Innate immune defense against malaria infection in the mosquito. Curr Opin Immunol 13:79–88

    Article  PubMed  CAS  Google Scholar 

  13. Osta MA, Christophides GK, Vlachou D, Kafatos FC (2004) Innate immunity in the malaria vector Anopheles gambiae: comparative and functional genomics. J Exp Biol 207:2551–2563

    Article  PubMed  CAS  Google Scholar 

  14. Jiang H, Kanost MR (2000) The clip-domain family of serine proteinases in arthropods. Insect Biochem Mol Biol 30:95–105

    Article  PubMed  CAS  Google Scholar 

  15. Kanost MR, Jiang H, Wang Y, Yu XQ, Ma C, Zhu Y (2001) Hemolymph proteinases in immune responses of Manduca sexta. Adv Exp Med Biol 484:319–328

    PubMed  CAS  Google Scholar 

  16. De Gregorio E, Spellman PT, Rubin GM, Lemaitre B (2001) Genome-wide analysis of the Drosophila immune response by using oligonucleotide microarrays. Proc Natl Acad Sci USA 98:12590–12595

    Article  PubMed  Google Scholar 

  17. Ligoxygakis P, Pelte N, Hoffmann JA, Reichhart JM (2002) Activation of Drosophila Toll during fungal infection by a blood serine protease. Science 297:114–116

    Article  PubMed  CAS  Google Scholar 

  18. Muta T, Hashimoto R, Miyata T, Nishimura H, Toh Y, Iwanaga S (1990) Proclotting enzyme from horseshoe crab hemocytes. cDNA cloning, disulfide locations, and subcellular localisation. J Biol Chem 265:22426–22433

    PubMed  CAS  Google Scholar 

  19. Muta T, Oda T, Iwanaga S (1993) Horseshoe crab coagulation factor B. A unique serine protease zymogen activated by cleavage of an Ile-Ile bond. J Biol Chem 268:21384–21388

    PubMed  CAS  Google Scholar 

  20. Dohke K (1973a) Studies on prophenoloxidase-activating enzyme from cuticle of the silkworm Bombyx mori. I. Activation reaction by the enzyme. Arch Biochem Biophys 157:203–209

    Article  PubMed  CAS  Google Scholar 

  21. Dohke K (1973b) Studies on prephenoloxidase-activating enzyme from cuticle of the silkworm Bombyx mori. II. Purification and characterization of the enzyme. Arch Biochem Biophys 157:210–221

    Article  PubMed  CAS  Google Scholar 

  22. Aspán A, Hall M, Söderhäll K (1990) The effect of endogenous proteinase inhibitors on the prophenoloxidase activating enzyme, a serine proteinase from crayfish haemocytes. Insect Biochem 20:485–492

    Article  Google Scholar 

  23. Aspán A, Söderhäll K (1991) Purification of prophenoloxidase from crayfish blood cells and its activation by an endogenous serine proteinase. Insect Biochem 21:363–373

    Article  Google Scholar 

  24. Chosa N, Fukumitsu T, Fujimoto K, Ohnishi E (1997) Activation of prophenoloxidase A1 by an activating enzyme in Drosophila melanogaster. Insect Biochem Mol Biol 27:61–68

    Article  PubMed  CAS  Google Scholar 

  25. Lee SY, Kwon TH, Hyun JH, Choi JS, Kawabata SI, Iwanaga S, Lee BL (1998a) In vitro activation of prophenoloxidase by two kinds of prophenoloxidase activating factors isolated from hemolymph of coleopteran, Holotrichia diomphalia larvae. Eur J Biochem 254:50–57

    Article  PubMed  CAS  Google Scholar 

  26. Lee SY, Cho MY, Hyun JH, Lee KM, Homma KI, Natori S, Kawabata SI, Iwanaga S, Lee BL (1998b) Molecular cloning of cDNA for pro-phenol-oxidase-activating factor I, a serine protease is induced by lipopolysaccharide or 1,3–glucan in coleopteran insect, Holotrichia diomphalia larvae. Eur J Biochem 257:615–621

    Article  PubMed  CAS  Google Scholar 

  27. Jiang HB, Wang Y, Kanost MR (1998) Pro-phenol oxidase activating proteinase from an insect, Manduca Sexta: A bacteria-inducible protein similar to Drosophila easter. Proc Natl Acad Sci 95:12220–12225

    Article  PubMed  CAS  Google Scholar 

  28. Zhu L, Song L, Zhao J, Li Ch, Chang Y (2006) Molecular cloning, characterization and expression of a serine proteinase with clip-domain homologue from scallop Chlamys farreri. Fish Shellfish Immunol doi:10.1016/j.fsi.2006.08.002

  29. Jacquinet E, Rao NV, Rao GV, Hoidal JR (2000) Cloning, genomic organization, chromosomal assignment and expression of a novel mosaic serine protease: epitheliasin. FEBS Lett 468:93–100

    Article  PubMed  CAS  Google Scholar 

  30. Ross J, Jiang H, Kanost MR, Wang Y (2003) Serine proteases and their homologs in the Drosophila melanogaster genome: an initial analysis of sequence conservation and phylogenetic relationship. Gene 304:117–131

    Article  PubMed  CAS  Google Scholar 

  31. Kumar SK, Tamura K, Jakobsen IB, Nei M (2000) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17:1244–1245

    Article  Google Scholar 

  32. Kawabata S, Muta T, Iwanaga S (1996) Clotting cascade and defense molecules found in the hemolymph of the horseshoe crab. In: Söderhäll K, Iwanaga S, Vasta G (eds) New directions in invertebrate immunology. Fair Haven, NJ, SOS publication, pp 255–283

    Google Scholar 

  33. Gaines PJ, Sampson CM, Rushlow KE, Stiegler GL (1999) Cloning of a family of serine protease genes from the cat flea Ctenocephalides felis. Insect Mol Biol 8:11–22

    Article  PubMed  CAS  Google Scholar 

  34. Yount NY, Yuan J, Tarver A, Castro T, Diamond G, Tran PA, Levy JN, McCullough C, Cullor JS, Bevins CL, Selsted ME (1999) Cloning and expression of bovine neutrophil β-defensins: biosynthetic profile during neutrophilic maturation and localization of mature peptide to novel cytoplasmic dense granules. J Biol Chem 274:26249–26258

    Article  PubMed  CAS  Google Scholar 

  35. Iwanaga S, Kawabata S, Muta T (1998) New types of clotting factors and defense molecules in horseshoe crab hemolymph: their structures and functions. J Biochem 123:1–15

    PubMed  CAS  Google Scholar 

  36. Dulinski R, Suder P, Guevara-Lora I, Rapala-Kozik M, Potempa J, Silberring J, Imamura T, Travis J, Kozik A (2003) Attenuated kinin release from human neutrophil-elastase pretreated kininogens by tissue and plasma kallikreins. Biol Chem 384:929–937

    Article  PubMed  CAS  Google Scholar 

  37. Bannach FG, Gutierrez A, Fowler BJ, Bugge TH, Degen JL, Parmer RJ, Miles LA (2002) Localization of regulatory elements mediating constitutive and cytokine-stimulated plasminogen gene expression. J Biol Chem 277:38579–38588

    Article  PubMed  CAS  Google Scholar 

  38. Dohke K (1973) Studies on prephenoloxidase-activating enzyme from cuticle of the silkworm Bombyx mori. II. Purification and characterization of the enzyme. Arch Biochem Biophys 157:210–221

    Article  PubMed  CAS  Google Scholar 

  39. Jiang H, Wang Y, Yu X-Q, Kanost MR (2003) Prophenoloxidase-activating proteinase-2 (PAP-2) from hemolymph of Manduca sexta: a bacteria-inducible serine proteinase containing two clip domains. J Biol Chem 278:3552–3561

    Article  PubMed  CAS  Google Scholar 

  40. Kopacek P, Grubnoffer L, Söderhäll K (1993) Isolation and. characterization of a hemagglutinin with affinity for lipoplysaccharides from. plasma of the crayfish Pacifastacus leniusculus. Dev Comp Immunol 17:407–418

    Article  PubMed  CAS  Google Scholar 

  41. Lavine MD, Strand MR (2002) Insect hemocytes and their role in immunity. Insect Biochem Mol Biol 32:1295–1309

    Article  PubMed  CAS  Google Scholar 

  42. Simser JA, Mulenga A, Macaluso KR, Azad AF (2004) An immune responsive factor D-like serine proteinase homologue identified from the American dog tick, Dermacentor variabilis. Insect Mol Biol 13:25–35

    Article  PubMed  CAS  Google Scholar 

  43. Jiang H, Wang Y, Kanost MR (1999) Four serine proteinases expressed in Manduca sexta haemocytes. Insect Mol Biol 8:39–53

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank all laboratory members for technical advice and helpful discussions. This research was supported by 863 High Technology Project (No. 2002AA626020) from the Chinese Ministry of Science and Technology, the grant (No. 40276045 and 30230280) from NSFC to Dr. Linsheng Song and Prof. Jianhai Xiang, and the grant (No.06SB011014) from South China Sea Institute of Oceanography, Chinese Academy of Sciences to Dr. Ling Zhu.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Ling Zhu or Linsheng Song.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhu, L., Song, L., Mao, Y. et al. A novel serine protease with clip domain from scallop Chlamys farreri . Mol Biol Rep 35, 257–264 (2008). https://doi.org/10.1007/s11033-007-9078-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-007-9078-2

Keywords

Navigation