Abstract
A C-type lectin-like protein was cloned and characterized from the Chinese shrimp Fenneropenaeus chinensis, and named as FcCTL. The results indicated that the full length cDNA of 859 bp had an open reading frame encoded a polypeptide of 220 amino acids with one carbohydrate-recognition domain, six conserved Cys and one key motif EPGD. The theoretical molecular weight and pI of mature protein was 25.3 kDa and 5.4. Sequence comparison of the deduced amino acid sequence of FcCTL showed varied identity of 26–34, 34, 31 and 30 % with those of F. chinensis, Portunus trituberculatus, Tetraodon nigroviridis, Penaeus monodon, respectively. qRT-PCR analysis indicated that FcCTL was expressed highest in hepatopancreas of normal shrimp, and it’s expression was up-regulated in hepatopancreas and gills post white spot syndrome virus challenge. The purified recombinant FcCTL showed higher antimicrobial activity against Gram-positive bacteria than against Gram-negative bacteria and fungi. And the hemagglutinating activity of rFcCTL could be completely inhibited by GlcNAc (5 μg/ml), LPS (2.5 μg/ml), d-galactose (100 mM) and maltose (100 mM). These data suggested that FcCTL might play an important role in shrimp immune and would be helpful to better understand the innate immunity mechanism of shrimp.
Similar content being viewed by others
References
Zhang Y, Qiu L, Song L (2009) Cloning and characterization of a novel C-type lectin gene from shrimp Litopenaeus vannamei. Fish Shellfish Immunol 26:183–192
Yu XQ, Gan H, Kanost MR (1999) Immulectin, an inducible C-type lectin from an insect, Manduca sexta, stimulates activation of plasma prophenol oxidase. Insect Biochem Mol Biol 29:585–597
Yu Y, Huang H, Feng K (2007) A short-form C-type lectin from amphioxus acts as a direct microbial killing protein via interaction with peptidoglycan and glucan. J Immunol 179:8425–8434
Odintsova NA, Belogortseva NI, Khomenko AV, Chikalovets IV, Luk’yanov PA (2001) Effect of lectin from the ascidian on the growth and the adhesion of HeLa cells. Mol Cell Biochem 221:133–138
Janeway CA, Medzhitov R (2002) Innate immune recognition. Annu Rev Immunol 20:197–216
Ma TH, Tiu SH, He JG, Chan SM (2007) Molecular cloning of a C-type lectin (LvLT) from the shrimp Litopenaeus vannamei: early gene down-regulation after WSSV infection. Fish Shellfish Immunol 23:430–437
Sun Y, Fu L, Jia Y (2008) A hepatopancreas-specific C-type lectin from the Chinese shrimp Fenneropenaeus chinensis exhibits antimicrobial activity. Mol Immunol 45:348–361
Robinson MJ, Sancho D, Slack EC, Leibundgut-Landmann S, Sousa CR (2006) Myeloid C-type lectins in innate immunity. Nat Immunol 7:1258–1265
Ezekowitz RA (2003) Role of the mannose-binding lectin in innate immunity. J Infect Dis 187(Suppl 2):S335–S339
Apostolopoulos V, McKenzie IF (2001) Role of the mannose receptor in the immune response. Curr Mol Med 1:469–474
Luo T, Yang H, Li F (2006) Purification, characterization and cDNA cloning of a novel lipopolysaccharide-binding lectin from the shrimp Penaeus monodon. Dev Comp Immunol 30:607–617
He JG, Deng M, Long QX, Zhou HM, Li GS, Yao P, Lu L, Chen YG, Weng SP, Miao SY, Jiang SG, Wu YY, Huang X, Zuo T, Yang XM, Yu N, Xie ST, Zhong W, Li GF, Ye QZ, Jiang JB, Chan SM, Mo F (2000) Theory and strategies for controlling white spot syndrome (WSS) of cultured Penaeus monodon in South China. Acta Sci Nat Univ Sunyatseni 39(Suppl):147–153
Lightner DV, Redman RM (1998) Shrimp diseases and current diagnostic methods. Aquaculture 164:201–220
Lo CF, Ho CH, Peng SE, Chen CH, Hsu HC, Chiu YL, Chang CF, Liu KF, Su MS, Wang CH, Kou GH (1996) White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp, crabs and other arthropods. Dis Aquat Organ 27:215–225
Wang X, Xu W, Zhang X (2009) A C-type lectin is involved in the innate immune response of Chinese white shrimp. Fish Shellfish Immunol 27:556–562
Wang X, Zhang X, Xu W (2009) A novel C-type lectin (FcLec4) facilitates the clearance of Vibrio anguillarum in vivo in Chinese white shrimp. Dev Comp Immunol 33:1039–1047
Liu Y, Li F, Dong B (2007) Molecular cloning, characterization and expression analysis of a putative C-type lectin (Fclectin) gene in Chinese shrimp Fenneropenaeus chinensis. Mol Immunol 44:598–607
Ma T, Benzie J, He J, Chan S (2008) PmLT, a C-type lectin specific to hepatopancreas is involved in the innate defense of the shrimp Penaeus monodon. J Invertebr Pathol 99:332–341
Song KK, Li DF, Zhang MC, Yang HJ, Ruan LW, Xu X (2010) Cloning and characterization of three novel WSSV recognizing lectins from shrimp Marsupenaeus japonicus. Fish Shellfish Immunol. doi:10.1016/j.fsi.2009.12.015
Lai XF, Kong J, Wang QY, Wang WJ, Meng XH (2011) Cloning and characterization of a β-1,3-glucan-binding protein from shrimp Fenneropenaeus chinensis. Mol Biol Rep 38:4527–4535
Lai XF, Kong J, Wang QY, Wang WJ (2010) Analysis of molecular basis related to WSSV-resistance in Fenneropenaeus chinensis by mRNA differential display PCR. J Fish Sci China 17(2):2–8
Amparyup P, Kondo H, Hirono I (2008) Molecular cloning, genomic organization and recombinant expression of a crustin-like antimicrobial peptide from black tiger shrimp Penaeus monodon. Mol Immunol 45:1085–1093
Zelensky AN, Gready JE (2005) The C-type lectin-like domain superfamily. FEBS J 272:6179–6217
Adams MD, Celniker SE, Holt RA (2000) The genome sequence of Drosophila melanogaster. Science 287:2185–2195
Schulenburg H, Hoeppner MP, Weiner J, Bornberg-Bauer E (2008) Specificity of the innate immune system and diversity of C-type lectin domain (CTLD) proteins in the nematode Caenorhabditis elegans. Immunobiology 213:237–250
Yu XQ, Kanost MR (2004) Immulectin-2, a pattern recognition receptor that stimulates hemocyte encapsulation and melanization in the tobacco hornworm Manduca sexta. Dev Comp Immunol 28:891–900
Yu XQ, Kanost MR (2000) Immulectin-2, a lipopolysaccharide-specific lectin from an insect, Manduca sexta, is induced in response to gram-negative bacteria. J Biol Chem 275:37373–37381
Yu XQ, Ma Y (2006) Calcium is not required for immulectin-2 binding, but protects the protein from proteinase digestion. Insect Biochem Mol Biol 36:505–516
Zheng P, Wang H, Zhao J (2008) A lectin (CfLec-2) aggregating Staphylococcus haemolyticus from scallop Chlamys farreri. Fish Shellfish Immunol 24:286–293
Drickamer K (1992) Engineering galactose-binding activity into a C-type mannose-binding protein. Nature 360:183–186
Tiffoche C, Chesnel A, Jego P, Le Pennec JP (1993) Isolation and characterization of a cDNA clone encoding a pleurodeles lectin. Eur J Biochem 213:901–907
Zhao ZY, Yin ZX, Xu XP (2009) A novel C-type lectin from the shrimp Litopenaeus vannamei possesses anti-white spot syndrome virus activity. J Virol 83:347–356
Zhang J, Li F, Wang Z, Xiang J (2007) Expression, purification, and characterization of recombinant Chinese shrimp crustin-like protein (CruFc) in Pichia pastoris. Biotechnol Lett 29:813–817
Supungul P, Tang S, Maneeruttanarungroj C, Rimphanitchayakit V, Hirono I, Aoki T, Tassanakajon A (2008) Cloning, expression and antimicrobial activity of crustinPm1, a major isoform of crustin, from the black tiger shrimp Penaeus monodon. Dev Comp Immunol 32(1):61–70
Acknowledgments
This work was financially supported by National High-Tech R&D Program of China (863 program, No. 2006AA10A406, No. 2010AA10A401) and by Jiangsu Key Laboratory of Marine Biotechnology program of China (No. 2008HS019).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lai, X., Kong, J., Wang, Q. et al. Identification and molecular characterization of a C-type lectin-like protein from Chinese shrimp (Fenneropenaeus chinensis). Mol Biol Rep 40, 2223–2230 (2013). https://doi.org/10.1007/s11033-012-2284-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-012-2284-6