Cite this paper:
XIA Dandan, MA Aijun, HUANG Zhihui, SHANG Xiaomei, CUI Wenxiao, YANG Zhi, QU Jiangbo. Molecular characterization and expression analysis of Lilytype lectin (SmLTL) in turbot Scophthalmus maximus, and its response to Vibrio anguillarum[J]. HaiyangYuHuZhao, 2018, 36(2): 508-518

Molecular characterization and expression analysis of Lilytype lectin (SmLTL) in turbot Scophthalmus maximus, and its response to Vibrio anguillarum

XIA Dandan1,2,3, MA Aijun1,3, HUANG Zhihui1,3, SHANG Xiaomei1,2,3, CUI Wenxiao1,2,3, YANG Zhi4, QU Jiangbo4
1 Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture;Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;
2 Ocean University of Shanghai, Shanghai 201306, China;
3 Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China;
4 Yantai Tianyuan Aquatic Limited Corporation, Yantai 264006, China
A full-length lily-type lectin (SmLTL) was identified from turbot (Scophthalmus maximus) in this study. By searching database for protein identification and function prediction, SmLTL were confirmed. The full-length cDNA of SmLTL is composed of 569 bp and contains a 339 bp ORF that encodes 112 amino acid residues. The SmLTL peptide is characterized by a specific β-prism architecture and contains three mannose binding sites in a three-fold internal repeat between amino acids 30-99; two of the repeats share the classical mannose binding domain (QxDxNxVxY) while the third binding site was similar to other fish-specific binding motifs (TxTxGxRxV). The primary, secondary, and tertiary structures of SmLTL were predicted and analyzed, indicating that the SmLTL protein was hydrophilic, contained 5.36% α-helices, 39.29% extended strands, 16.07% β-folds, and 39.29% random coils, and three β-folds. Quantitative realtime polymerase chain reaction (qPCR) analysis revealed that the SmLTL mRNA was abundantly expressed in skin, gill, and intestine. Low levels of SmLTL expression were observed in other tissues. The expression of SmLTL in gill, skin and intestine increased at mRNA level after stimulation of Vibrio anguillarum, our results suggest that SmLTL serve as the first line of defence against microbial infections and play a pivotal role in the innate mucosal immune system. The current study indicates that SmLTL is a member of the lilytype lectin family and the information reported here will provide an important foundation for future research on the role of this protein.
Key words:    lily-type lectin|full-length of cDNA|quantitative real-time polymerase chain reaction|protein structure|Vibrio anguillarum   
Received: 2016-10-03   Revised:
PDF (952 KB) Free
Print this page
Add to favorites
Email this article to others
Articles by XIA Dandan
Articles by MA Aijun
Articles by HUANG Zhihui
Articles by SHANG Xiaomei
Articles by CUI Wenxiao
Articles by YANG Zhi
Articles by QU Jiangbo
Afolabi-Balogun N B, Inuwa H M, Ishiyaku M F, Bakareodunoola M T, Nok A J. 2012. Isolation and characterization of a mannose-binding insecticidal lectin gene from Allium sativum (garlic) and its putative role in insect resistance using bioinformatics tools. Infect. Genet.Evol., 12(7):1 508-1 512.
Alexander J B, Ingram G A. 1992. Noncellular nonspecific defense mechanism of fish. Annual Review of Fish Diseases, 2:249-279.
Arasu A, Kumaresan V, Sathyamoorthi A, Palanisamy R, Prabha N, Bhatt P, Roy A, Thirumalai M K, Gnaname A J, Pasupuleti M, Marimuthu K, Arockiaraj J. 2013. Fish lily type lectin-1 contains β-prism architecture:immunological characterization. Mol. Immunol., 56(4):497-506.
Barre A, Bourne Y, Van Damme E J M, Peumans W J, Rougé P. 2001. Mannose-binding plant lectins:different structural scaffolds for a common sugar-recognition process. Biochimie, 83(7):645-651.
Berthelot C, Brunet F, Chalopin D, Juanchich A, Bernard M, Noël B, Bento P, Da Silva C, Labadie K, Alberti A, Aury J M, Louis A, Dehais P, Bardou P, Montfort J, Klopp C, Cabau C, Gaspin C, Thorgaard G H, Boussaha M, Quillet E, Guyomard R, Galiana D, Bobe J, Volff J N, Genêt C, Wincker P, Jaillon O, Crollius H R, Guiguen Y. 2014. The rainbow trout genome provides novel insights into evolution after whole-genome duplication in vertebrates.Nat. Commun., 5:3 657.
Bhat G G, Shetty K N, Nagre N N, Neekhra V V, Lingaraju S, Bhat R S, Inamdar S R, Suguna K, Swamy B M. 2010. Purification, characterization and molecular cloning of a monocot mannose-binding lectin from Remusatia vivipara with nematicidal activity. Glycoconj. J., 27(3):309-320.
Chandra N R, Ramachandraiah G, Bachhawat K, Dam T K, Surolia A, Vijayan M. 1999. Crystal structure of a dimeric mannose-specific agglutinin from garlic:quaternary association and carbohydrate specificity. J. Mol. Biol., 285(3):1 157-1 168.
Chen S R., Tang J X, Cheng J M, Li J, Jin C, Li X Y, Deng S L, Zhang Y, Wang X X, Liu Y X. 2015. Loss of gata4 in sertoli cells impairs the spermatogonial stem cell niche and causes germ cell exhaustion by attenuating chemokine signaling. Oncotarget, 6(35):37 012-37 027.
Chen Y D, Zhou S H, Jiang Z Q, Wang X L, Liu Y. 2016.Chemokine receptor CXCR3 in turbot (Scophthalmus maximus):cloning, characterization and its responses to lipopolysaccharide. Fish Physiol. Biochem., 42(2):659-671.
Chi H, Hu Y H. 2016. Stomatin-like protein 2 of turbot Scopthalmus maximus:gene cloning, expression profiling and immunoregulatory properties. Fish & Shellfish Immun., 49:436.
Ding J J, Bao J K, Zhu D Y, Zhang Y, Wang D C. 2008.Crystallization and preliminary x-ray diffraction analysis of a novel mannose-binding lectin with antiretroviral properties from Polygonatum cyrtonema hua. Protein Pept. Lett., 15(4):411-414.
Dodd R B, Drickamer K. 2001. Lectin-like proteins in model organisms:implications for evolution of carbohydratebinding activity. Glycobiology, 11(5):71R-79R.
Gao C B, Fu Q, Zhou S, Song L, RenY C, Dong X Y, Su B F, Li C. 2016. The mucosal expression signatures of g-type lysozyme in turbot (Scophthalmus maximus) following bacterial challenge. Fish Shellfish Immunol., 54:612-619.
Goel C, Barat A, Pande V, Sahoo P K. 2015. Molecular cloning and characterization of mannose binding lectin homologue from snow trout (Schizothorax richardsonii). The protein J., 34(1):1-8.
Goto-Nance R, Watanabe Y, Kamiya H, Ida H. 1995.Characterization of lectins feom the skin mucus of the Lonach Misgurnus anguillicaudatus. Fish. Sci., 61(1):137-140.
Guardiola F A, Cuesta A, Arizcun M, Meseguer J, Esteban M A. 2014. Comparative skin mucus and serum humoral defence mechanisms in the teleost gilthead seabream(Sparus aurata). Fish Shellfish Immunol., 36(2):545-551.
Hester G, Wright C S. 1996. The mannose-specific bulb lectin from Galanthus nivalis (snowdrop) binds mono-and dimannosides at distinct sites. Structure analysis of refined complexes at 2.3 Å and 3.0 Å resolution. J. Mol. Biol., 262(4):516-531.
Hoffmann J A, Kafatos F C, Janeway C A, Ezekowitz R A B. 1999. Phylogenetic perspectives in innate immunity.Science, 284(5418):1 313-1 318.
Huang M M, Song X Y, Zhao J M, Mu C K, Wang L L, Zhang H, Zhou Z, Liu X L, Song L S. 2013. A C-type lectin(AiCTL-3) from bay scallop Argopecten irradians with mannose/galactose binding ability to bind various bacteria. Gene, 531(1):31-38.
Huang Z H, Ma A J, Wang X A. 2011. The immune response of turbot, Scophthalmus maximus (L.), skin to high water temperature. J. Fish Dis., 34(8):619-627.
Ingram G A. 1980. Substances involved in the natural resistance of fish to infection-a review. J. Fish Biol., 16(1):23-60.
Kai G Y, Zhao L X, Zheng J G, Zhang L, Miao Z Q, Sun X F, Tang K X. 2004. Isolation and characterization of a new mannose-binding lectin gene from Taxus media. J. Biosci., 29(4):399-407.
Kamiya H, Muramoto K, Goto R. 1988. Purification and properties of agglutinins from conger eel, Conger myriaster (Brevoort), skin mucus. Dev. Comp. Immunol., 12(2):309-318.
Ke J Y, Chen Y S, Rao X Z. 2005. Lectins and its biological function. J. Ningde Teach. Coll. Nat. Sci., 17(1):19-22.(in Chinese with English abstract)
Khong H K, Kuah M K, Jaya-Ram A, Shu-Chien A C. 2009.Prolactin receptor mRNA is upregulated in discus fish(Symphysodon aequifasciata) skin during parental phase.Comp. Biochem. Physiol. B:Biochem. Mol. Biol., 153:18-28.
Kim B S, Nam B H, Kim J W, Park H J, Song J H, Park C I. 2011. Molecular characterisation and expression analysis of a fish-egg lectin in rock bream, and its response to bacterial or viral infection. Fish & Shellfish Immun., 31(31):1 201-1 207.
Kyte J, Doolittle R F. 1982. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol., 157(1):105-132.
Leong J S, Jantzen S G, von Schalburg K R, Cooper G A, Messmer A M, Liao N Y, Munro s, Moore R, Holt R A, Jones S J M, Davidson W S, Koop B F. 2010. Salmo salar and Esox lucius full-length cDNA sequences reveal changes in evolutionary pressures on a posttetraploidization genome. BMC Genomics, 11(1):279.
Lin J Y, Hu G B, Yu C H, Li S, Liu Q M, Zhang S C. 2015.Molecular cloning and expression studies of the adapter molecule myeloid differentiation factor 88 (MyD88) in turbot (Scophthalmus maximus). Dev. Comp. Immunol., 52(2):166-171.
Liu F Q, Su B F, Gao C B, Zhou S, Song L, Tan F H, Dong X Y, Ren Y C, Li C. 2016. Identification and expression analysis of TLR2 in mucosal tissues of turbot(Scophthalmus maximus L.) following bacterial challenge. Fish Shellfish Immunol., 55:654-661.
Luo Y, Xu X, Liu J, Li J, Sun Y, Liu Z, Liu J Z, Van Damme E, Balzarini J, Bao J. 2007. A novel mannose-binding tuber lectin from Typhonium divaricatum (L.) Decne (family Araceae) with antiviral activity against HSV-Ⅱ and antiproliferative effect on human cancer cell lines. BMB Rep., 40(3):358-367.
Ma A J, Guo J L, Wang X A, Huang Z H. Wang T, Shang X M. 2014. Family selection and estimation of disease resistance in turbot, Scophthalmus maximus. J. Fish. Sci.China, 21(3):484-493. (in Chinese with English abstract)
Ma A J, Huang Z H, Wang X A. 2013. Changes in protein composition of epidermal mucus in turbot Scophthalmus maximus (L.) under high water temperature. Fish Physiol.Biochem., 39(6):1 411-1 418.
Meng Y Q, Liu X F, Liu Y, Chang X Q, Wang X L, Jiang Z Q. 2013. Chemokine receptor genes CCR3 and CCR9 in turbot (Scophthalmus maximus):cloning and tissue distribution. J. Fish. Sci. China, 20(5):918-930. (in Chinese with English abstract)
Moore J D, Ototake M, Nakanishi T. 1998. Particulate antigen uptake during immersion immunisation of fish:the effectiveness of prolonged exposure and the roles of skin and gill. Fish Shellfish Immunol., 8(6):393-408.
Park H J, Jeong J M, Bae J S, Kim J W, An C M, Min B H, Kim S Y, Myeong J I, Hwang H K, Park C I. 2016. Molecular cloning and expression analysis of a new lily-type lectin in the rock bream, Oplegnathus fasciatus. Dev. Comp.Immunol., 65:25-30.
Pereira P R, Winter H C, Verícimo M A, Meagher J L, Stuckey J A., Goldstein I J, Paschoalin V M F, Silva J T. 2014. Structural analysis and binding properties of isoforms of tarin, the gna-related lectin from Colocasia esculenta.BBA-Proteins proteome., 1854(1):20-30.
Sharon N, Lis H. 2004. History of lectins:from hemagglutinins to biological recognition molecules. Glycobiology, 14(11):53R-62R.
Shephard K L. 1994. Functions for fish mucus. Rev. Fish Biol.Fish., 4(4):401-429.
Shiomi K, Uematsu H, Ito H, Yamanaka H, Kikuchi T. 1990.Purification and properties of a lectin in the skin mucus of the dragonet Repomucenus richardsonii. Nippon Suisan Gakk., 56(1):119-123.
Shiomi K, Uematsu H, Yamanaka H, Kikuchi T. 1989.Purificatioin and characterization of a galactose-binding lectin from the skin mucus of the conger eel Conger myriaster. Comp. Biochem. Physiol. B Comp. Biochem., 92(2):255-261.
Smeets K, Van Damme E J M, Verhaert P, Barre A, Rougé P, Van Leuven F, Peumans W J. 1997. Isolation, characterization and molecular cloning of the mannosebinding lectins from leaves and roots of garlic (Allium sativum L.). Plant Mol. Biol., 33(2):223-234.
Spitzer R H, Koch E A. 1998. Hagfish skin and slime glands.In:Jørgensen J M, Lomholt J P, Weber R E, Malte H eds.The Biology of Hagfishes. Springer, Netherlands. p.109-132.
Suzuki Y, Tasumi S, Tsutsui S, Okamoto M, Suetake H. 2003.Molecular diversity of skin mucus lectins in fish. Comp.Biochem. Physiol. B Biochem. Mol. Biol., 136(4):723-730.
Tasumi S, Ohira T, Kawazoe I, Suetake H, Suzuki Y, Aida K. 2002. Primary structure and characteristics of a lectin from skin mucus of the Japanese eel Anguilla japonica. J.Biol. Chem., 277(30):27 305-27 311.
Tian Q, Wang W, Miao C, Peng H, Liu B, Leng F, Dai L, Chen F, Bao J. 2008. Purification, characterization and molecular cloning of a novel mannose-binding lectin from rhizomes of Ophiopogon japonicus, with antiviral and antifungal activities. Plant Sci., 175(6):877-884.
Toda M, Goto-Nance R, Muramoto K, Kamiya H. 1996.Characterization of the lectin from the skin mucus of the Kingklip Genypterus capensis. Fish. Sci., 62(1):138-141.
Toranzo A E, Santos Y, Barja J L. 1997. Immunization with bacterial antigens:vibrio infections. Dev. Biol. Stand., 90:93-105.
Tsutsui S, Tasumi S, Suetake H, Suzuki Y. 2003. Lectins homologous to those of monocotyledonous plants in the skin mucus and intestine of pufferfish, Fugu rubripes. J.Biol. Chem., 278(23):20 882-20 889.
Tsutsui S, Tasumi S, Suetake H, Kikuchi K, Suzuki Y. 2006.Carbohydrate-binding site of a novel mannose-specific lectin from fugu (Takifugu rubripes) skin mucus. Comp.Biochem. Physiol. B Biochem. Mol. Biol., 143(4):514-519.
van Damme E J M, De Clercq N, Claessens F, Hemschoote K, Peeters B, Peumans W J. 1991. Molecular cloning and characterization of multiple isoforms of the snowdrop(Galanthus nivalis L.) lectin. Planta, 186(1):35-43.
van Damme E J M, Smeets K, Engelborghs I, Aelbers H, Balzarini J, Pusztai A, van Leuven F, Goldstein I J, Peumans W J. 1993. Cloning and characterization of the lectin cDNA clones from onion, shallot and leek. Plant Mol. Biol., 23(2):365-376.
van Damme E J M, Smeets K, Van Leuven F, Peumans W J. 1994. Molecular cloning of mannose-binding lectins from Clivia miniata. Plant Mol. Boil., 24(5):825-830.
van der Marel M, Caspari N, Neuhaus H, Meyer W, Enss M L, Steinhagen D. 2010. Changes in skin mucus of common carp, Cyprinus carpio L., after exposure to water with a high bacterial load. J. Fish Dis., 33(5):431-439.
Vasta G R, Ahmed H, Odom E W. 2004. Structural and functional diversity of lectin repertoires in invertebrates, protochordates and ectothermic vertebrates. Curr. Opin.Struct. Biol., 14(5):617-630.
Wang X W, Zhang X W, Xu W T, Zhao X F, Wang J X. 2009.A novel C-type lectin (FcLec4) facilitates the clearance of Vibrio anguillarum in vivo in Chinese white shrimp. Dev.Comp. Immunol., 33(9):1 039-1 047.
Zhao X Y, Yao J H, Liao Z H, Zhang H Y, Chen F, Wang L, Lu Y Q, Sun X F, Yu S Q, Tang K X. 2003. Molecular cloning of a novel mannose-binding lectin gene from Arisaema heterophyllum. Plant Sci., 165(1):55-60.