Characterization and comparison of two C-type lectins with novel key motifs from mud crab <i>Scylla paramamosain</i> -- Journal of Oceanology and Limnology,2022,40(4):1628-1639

	Cite this paper:
	Boxin ZENG, Taiwei DONG, Yanting XIA, Shun YANG, Mengmeng HUANG, Hui FEI. Characterization and comparison of two C-type lectins with novel key motifs from mud crab Scylla paramamosain[J]. Journal of Oceanology and Limnology, 2022, 40(4): 1628-1639

Characterization and comparison of two C-type lectins with novel key motifs from mud crab Scylla paramamosain

Boxin ZENG¹, Taiwei DONG¹, Yanting XIA¹, Shun YANG^1,2, Mengmeng HUANG^1,2, Hui FEI^1,2

1 College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China;
2 Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, Zhejiang Sci-Tech University, Hangzhou 310018, China

Abstract:

As an important pattern recognition receptor (PRR) in the innate immune system, C-type lectin plays an important role in the innate immune process of invertebrates. Two C-type lectins SpCTL-C and SpCTL-D were characterized from mud crab (Scylla paramamosain). The predicted SpCTL-C and SpCTL-D proteins both contain a single carbohydrate-recognition domain (CRD) with key motif GlnPro-Ala (QPA) and Met-Pro-Ala (MPA), respectively. SpCTL-C and SpCTL-D transcripts distributed in all examined tissues, and the expression level was the highest in hepatopancreas. As PRR, the purified recombinant proteins rSpCTL-C and rSpCTL-D have high affinity for three kinds of pathogen-associated molecular patterns (PAMPs):β-glucan, lipopolysaccharide, and peptidoglycan. Besides, rSpCTL-D can bind to all nine microorganisms tested, while rSpCTL-C can bind to seven microorganisms except for Staphylococcus aureus and Micrococcus luteus. Both rSpCTL-C and rSpCTL-D showed agglutination activity towards fungi Pichia pastoris and Saccharomyces cerevisiae. However, rSpCTL-C and rSpCTL-D exhibited different antimicrobial activities:rSpCTL-D has a certain inhibitory effect on the growth of Vibrio fluvialis and M. luteus, while rSpCTL-C has no obvious inhibitory activity. The results show that rSpCTL-C and rSpCTL-D had better phagocytosis-promoting effect on M. luteus than the negative control. Meanwhile, both rSpCTL-C and rSpCTL-D had certain encapsulation-promoting activity. Collectively, two C-type lectins with novel key motifs make an important impact as PRR in immune response towards pathogens. At the same time, they play different functions in the innate immunity of mud crab S. paramamosain.

Key words: Scylla paramamosain|immune response|innate immunity|C-type lectin

Received: 2021-06-09 Revised:

Tools

PDF (1933 KB) Free

Print this page

Add to favorites

Email this article to others

Authors

Articles by Boxin ZENG

Articles by Taiwei DONG

Articles by Yanting XIA

Articles by Shun YANG

Articles by Mengmeng HUANG

Articles by Hui FEI

References:
Alenton R R R, Koiwai K, Miyaguchi K, Kondo H, Hirono I.2017.Pathogen recognition of a novel C-type lectin from Marsupenaeus japonicus reveals the divergent sugarbinding specificity of QAP motif.Scientific Reports, 7:45818, https://doi.org/10.1038/srep45818.
Brubaker S W, Bonham K S, Zanoni I, Kagan J C.2015.Innate immune pattern recognition:a cell biological perspective.Annual Review of Immunology, 33(1):257-290, https://doi.org/10.1146/annurev-immunol-032414-112240.
Cambi A, Koopman M, Figdor C G.2005.How C-type lectins detect pathogens.Cellular Microbiology, 7(4):481-488, http://doi.org/10.1111/j.1462-5822.2005.00506.x.
Cao J S, Lin Z Y, Zhang Y L, Zhang X Y, Li S K, Zhang N, Zou W H, Li Y Y, Chen J H, Wang X Y.2013.Cloning and characterization of the SpLRR cDNA from green mud crab, Scylla paramamosain.Fish & Shellfish Immunology, 34(1):129-135, https://doi.org/10.1016/j.fsi.2012.10.018.
Christophides G K, Zdobnov E, Barillas-Mury C, Birney E, Blandin S, Blass C, Brey P T, Collins F H, Danielli A, Dimopoulos G, Hetru C, Hoa N T, Hoffmann J A, Kanzok S M, Letunic I, Levashina E A, Loukeris T G, Lycett G, Meister S, Michel K, Moita L F, Mü ller H M, Osta M A, Paskewitz S M, Reichhart J M, Rzhetsky A, Troxler L, Vernick K D, Vlachou D, Volz J, Mering C V, Xu J N, Zheng L B, Bork P, Kafatos F C.2002.Immunityrelated genes and gene families in Anopheles gambiae.Science, 298(5591):159-165, https://doi.org/10.1126/science.1077136.
Coteur G, Warnau M, Jangoux M, Dubois P.2002.Reactive oxygen species (ROS) production by amoebocytes of Asterias rubens (Echinodermata).Fish & Shellfish Immunology, 12(3):187-200, https://doi.org/10.1006/fsim.2001.0366.
Drickamer K, Taylor M E.1993.Biology of animal lectins.Annual Review of Cell Biology, 9(1):237-264, https://doi.org/10.1146/annurev.cb.09.110193.001321.
Drickamer K.1993.Evolution of Ca²⁺-dependent animal lectins.Progress in Nucleic Acid Research and Molecular Biology, 45:207-232, https://doi.org/10.1016/S0079-6603(08)60870-3.
Drickamer K.1999.C-type lectin-like domains.Current Opinion in Structural Biology, 9(5):585-590, https://doi.org/10.1016/S0959-440X (99)00009-3.
Huang M M, Mu C K, Wu Y H, Ye F, Wang D, Sun C, Lv Z B, Han B N, Wang C L, Xu X W.2017.The functional characterization and comparison of two single CRD containing C-type lectins with novel and typical key motifs from Portunus trituberculatus.Fish & Shellfish Immunology, 70:398-407, https://doi.org/10.1016/j.fsi.2017.09.029.
Kang T, Xia Y T, Dong T W, Zheng X Y, Yang S, Qian S C, Huang M M, Fei H.2021.C-type lectin with a QPN motif from swimming crab Portunus trituberculatus displays broad nonself-recognition ability and functions as an opsonin.Developmental and Comparative Immunology, 120:104066, https://doi.org/10.1016/j.dci.2021.104066.
Lan T Y, Li Z, Peng M X, Niu D H, Li Y, Li J L.2020.A fourCRD C-type lectin from razor clam Sinonovacula constricta mediates agglutination and phagocytosis.Gene, 728:144287, https://doi.org/10.1016/j.gene.2019.144287.
Ling E J, Yu X Q.2006.Cellular encapsulation and melanization are enhanced by immulectins, pattern recognition receptors from the tobacco hornworm Manduca sexta.Developmental & Comparative Immunology, 30(3):289-299, https://doi.org/10.1016/j.dci.2005.05.005.
Liu Y C, Li F H, Dong B, Wang B, Luan W, Zhang X J, Zhang L S, Xiang J H.2007.Molecular cloning, characterization and expression analysis of a putative C-type lectin(Fclectin) gene in Chinese shrimp Fenneropenaeus chinensis.Molecular Immunology, 44(4):598-607, https://doi.org/10.1016/j.molimm.2006.01.015.
Loker E S, Adema C M, Zhang S M, Kepler T B.2004.Invertebrate immune systems-not homogeneous, not simple, not well understood.Immunological Reviews, 198(1):10-24, https://doi.org/10.1111/j.0105-2896.2004.0117.x.
Patel K D, Cuvelier S L, Wiehler S.2002.Selectins:critical mediators of leukocyte recruitment.Seminars in Immunology, 14(2):73-81, http://doi.org/10.1006/smim.2001.0344.
Rowley A F, Powell A.2007.Invertebrate immune systems specific, quasi-specific, or nonspecific? The Journal of immunology, 179(11):7209-7214, https://doi.org/10.4049/jimmunol.179.11.7209.
Soares-Da-Silva I M, Ribeiro J, Valongo C, Pinto R, Vilanova M, Bleher R, Machado J.2002.Cytometric, morphologic and enzymatic characterisation of haemocytes in Anodonta cygnea.Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology, 132(3):541-553, https://doi.org/10.1016/S1095-6433(02)00039-9.
Taylor M E.1993.Carbohydrate-recognition proteins of macrophages and related cells.In:Horton M A ed.Macrophages and Related Cells.Blood Cell Biochemistry, Vol.5.Springer, Boston, MA.p.347-370.
Tran N T, Kong T T, Zhang M, Li S K.2020.Pattern recognition receptors and their roles on the innate immune system of mud crab (Scylla paramamosain).Developmental & Comparative Immunology, 102:103469, https://doi.org/10.1016/j.dci.2019.103469.
Wang G Y, Lei Y T, Kang T, Li Z, Fei H, Zeng B X, Zhou P, Wang C S, Lv Z B, Huang M M, Xu X W.2020.Two C-type lectins (ReCTL-1, ReCTL-2) from Rimicaris exoculata display broad nonself recognition spectrum with novel carbohydrate binding specificity.Fish & Shellfish Immunology, 96:152-160, https://doi.org/10.1016/j.fsi.2019.11.068.
Wang X W, Gao J, Xu Y H, Xu J D, Fan Z X, Zhao X F, Wang J X.2017.Novel pattern recognition receptor protects shrimp by preventing bacterial colonization and promoting phagocytosis.The Journal of Immunology, 198(8):3045-3057, https://doi.org/10.4049/jimmunol.1602002.
Wang X W, Xu J D, Zhao X F, Vasta G R, Wang J X.2014.A shrimp C-type lectin inhibits proliferation of the hemolymph microbiota by maintaining the expression of antimicrobial peptides.Journal of Biological Chemistry, 289(17):11779-11790, https://doi.org/10.1074/jbc.M114.552307.
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.Developmental & Comparative Immunology, 33(9):1039-1047, https://doi.org/10.1016/j.dci.2009.05.004.
Wei X Y, Wang L M, Sun W W, Zhang M, Ma H Y, Zhang Y L, Zhang X X, Li S K.2018.C-type lectin B (SpCTL-B) regulates the expression of antimicrobial peptides and promotes phagocytosis in mud crab Scylla paramamosain.Developmental and Comparative Immunology, 84:213-229, https://doi.org/10.1016/j.dci.2018.02.016.
Weis W I, Taylor M E, Drickamer K.1998.The C-type lectin superfamily in the immune system.Immunological Reviews, 163(1):19-34, https://doi.org/10.1111/j.1600-065X.1998.tb01185.x.
Yu Y H, Yu Y C, Huang H Q, Feng K X, Pan M M, Yuan S C, Huang S F, Wu T, Guo L, Dong M L, Chen S W, Xu A L.2007.A short-form C-type lectin from amphioxus acts as a direct microbial killing protein via interaction with peptidoglycan and glucan.The Journal of Immunology, 179(12):8425-8434, https://doi.org/10.4049/jimmunol.179.12.8425.
Zelensky A N, Gready J E.2003.Comparative analysis of structural properties of the C-type-lectin-like domain(CTLD).Proteins:Structure, Function, and Bioinformatics, 52(3):466-477, https://doi.org/10.1002/prot.10626.
Zelensky A N, Gready J E.2005.The C-type lectin-like domain superfamily.The FEBS Journal, 272(24):6179-6217, https://doi.org/10.1111/j.1742-4658.2005.05031.x.
Zhang W J, Zhang Z Y, Mu C K, Li R H, Ye Y F, Zhang H, Song W W, Shi C, Liu L, Wang C L.2019.Molecular cloning, characterization, and expression of a C-type lectin from Scylla paramamosain, which might be involved in the innate immune response.Fish & Shellfish Immunology, 93:251-257, https://doi.org/10.1016/j.fsi.2019.07.035.
Zhu Y T, Zhang X, Wang S C, Li W W, Wang Q.2016.Antimicrobial functions of EsLecH, a C-type lectin, via JNK pathway in the Chinese mitten crab, Eriocheir sinensis.Developmental & Comparative Immunology, 61:225-235, https://doi.org/10.1016/j.dci.2016.04.007.