镰形扇头蜱卵巢中丝氨酸蛋白酶抑制分子群的克隆和生物信息学分析
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摘要
丝氨酸蛋白酶抑制分子(serine protease inhibitor,Serpin)参与多种生物学过程,对蜱的吸血发育、生殖产卵和先天免疫系统反应等具有重要作用,在蜱体内以多分子形式存在。为了研究蜱生殖相关的Serpin分子群,本研究通过构建镰形扇头蜱(Rhipicephalus haemaphysaloides)卵巢转录组测序数据库,发现并克隆了10条Serpin家族基因(RhS),其中3条为分泌型Serpin,6条为细胞内Serpin和1条亲缘关系较远的Serpin。本研究通过生物信息学方法分析其理化性质、遗传进化关系,预测其结构和功能,为镰形扇头蜱Serpin分子群的功能解析和应用研究提供了重要基础。
Serine protease inhibitor(Serpin) participates in a variety of biological functions and plays an important role in blood meal and development, reproductive spawning and innate immune system responses of ticks, and exists in multi-molecule form in ticks. In order to study the population of Serpin related to tick reproduction, 10 Serpin family genes(RhS) were found and cloned by ovarian transcriptome sequencing database of Rhipicephalus haemaphysaloides, of which 3 were secreted Serpin, 6 were intracellular Serpin and a Serpin with a distant relationship. Bioinformatics analysis of its physical and chemical properties, genetic evolution relationship, predict its structure and function, provide important basis for functional analysis and application of Serpin molecular cluster of Rhipicephalus haemaphysaloides.
Serine protease inhibitor(Serpin) participates in a variety of biological functions and plays an important role in blood meal and development, reproductive spawning and innate immune system responses of ticks, and exists in multi-molecule form in ticks. In order to study the population of Serpin related to tick reproduction, 10 Serpin family genes(RhS) were found and cloned by ovarian transcriptome sequencing database of Rhipicephalus haemaphysaloides, of which 3 were secreted Serpin, 6 were intracellular Serpin and a Serpin with a distant relationship. Bioinformatics analysis of its physical and chemical properties, genetic evolution relationship, predict its structure and function, provide important basis for functional analysis and application of Serpin molecular cluster of Rhipicephalus haemaphysaloides.
引文
[1]Huntington J A.Serpin structure,function and dysfunction[J].J Thromb Haemost,2011,9(suppl 1):26-34.
[2]van Gent D,Sharp P,Morgan K,et al.Serpins:structure,function and molecular evolution[J].Int J Biochem Cell Biol,2003,35(11):1536-1547.
[3]Reichhart J M.Tip of another iceberg:drosophila serpins[J].Trends Cell Biol,2005,15(12):659-665.
[4]Meekins D A,Kanost M R,Michel K.Serpins in arthropod biology[J].Semin Cell Dev Biol,2017,62:105-119.
[5]Law R H,Zhang Q,McGowan S,et al.An overview of the serpin superfamily[J].Genome Biol,2006,7(5):216.
[6]Irving J A,Pike R N,Lesk A M,et al.Phylogeny of the serpin superfamily:implications of patterns of amino acid conservation for structure and function[J].Genome Res,2000,10(12):1845-1864.
[7]Gulley M M,Zhang X,Michel K.The roles of serpins in mosquito immunology and physiology[J].J Insect Physiol,2013,59(2):138-147.
[8]PáleníkováJ,LieskovskáJ,LanghansováH,et al.Ixodes ricinus salivary serpin IRS-2 affects Th17differentiation via inhibition of the interleukin-6/STAT-3 signaling pathway[J].Infect Immun,2015,83(5):1949-1956.
[9]赵丽芳,陶美林,潘国庆.丝氨酸蛋白酶抑制剂超家族的研究进展[J].蚕业科学,2016,42(03):532-540.
[10]Rau J C,Beaulieu L M,Huntington J A,et al.Serpins in thrombosis,hemostasis and fibrinolysis[J].J Thromb Haemost.2007,5(suppl1):102-115.
[11]Francischetti I M,Sa-Nunes A,Mans B J,et al.The role of saliva in tick feeding[J].Front Biosci(Landmark Ed),2009,14:2051-2088.
[12]Leboulle G,Crippa M,Decrem Y,et al.Characterization of a novel salivary immunosuppressive protein from Ixodes ricinus ticks[J].J Biol Chem,2002,277(12):10083-10089.
[13]Prevot P P,Adam B,Boudjeltia K Z,et al.Antihemostatic effects of a serpin from the saliva of the tick Ixodes ricinus[J].J Biol Chem,2006,281(36):26361-26369.
[14]Anderson J M,Sonenshine D E,Valenzuela J G.Exploring the mialome of ticks:an annotated catalogue of midgut transcripts from the hard tick,Dermacentor variabilis(Acari:Ixodidae)[J].BMC Genomics,2008,9:552.
[15]Pak S C,Kumar V,Tsu C,et al.SRP-2 is a cross-class inhibitor that participates in postembryonic development of the nematode Caenorhabditis elegans:initial characterization of the clade L serpins[J].J Biol Chem,2004,279(15):15448-15459.
[16]Rodriguez-Valle M,Vance M,Moolhuijzen P M,et al.Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3(RMS-3)[J].Ticks Tick Borne Dis,2012,3(3):159-169.
[17]邓振,何庆丰,彭礼飞.中性粒细胞丝氨酸蛋白酶及其抑制剂与炎症[J].中国药理学通报,2014,30(3):314-317.
[18]Bode W.The structure of thrombin:a janus-headed proteinase[J].Semin Thromb Hemost,2006,32(Suppl 1):16-31.
[19]Prevot P P,Couvreur B,Denis V,et al.Protective immunity against Ixodes ricinus induced by a salivary serpin[J].Vaccine,2007,25(17):3284-3292.
[20]Prevot P P,Beschin A,Lins L,et al.Exosites mediate the anti-inflammatory effects of a multifunctional serpin from the saliva of the tick Ixodes ricinus[J].FEBS J,2009,276(12):3235-3246.
[21]Abraham E G,Pinto S B,Ghosh A,et al.An immuneresponsive serpin,SRPN6,mediates mosquito defense against malaria parasites[J].Proc Natl Acad Sci USA,2005,102(45):16327-16332.
[22]Pinto S B,Kafatos F C,Michel K.The parasite invasion marker SRPN6 reduces sporozoite numbers in salivary glands of Anopheles gambiae[J].Cell Microbiol,2008,10(4):891-898.
[23]Luo L M,Zhao L,Wen H L,et al.Haemaphysalis longicornis ticks as reservoir and vector of severe fever with Thrombocytopenia Syndrome Virus in China[J].Emerg Infect Dis,2015,21(10):1770-1776.
[24]Yu Y,Cao J,Zhou Y,et al.Isolation and characterization of two novel serpins from the tick Rhipicephalus haemaphysaloides[J].Ticks Tick Borne Dis,2013,4(4):297-303.
[2]van Gent D,Sharp P,Morgan K,et al.Serpins:structure,function and molecular evolution[J].Int J Biochem Cell Biol,2003,35(11):1536-1547.
[3]Reichhart J M.Tip of another iceberg:drosophila serpins[J].Trends Cell Biol,2005,15(12):659-665.
[4]Meekins D A,Kanost M R,Michel K.Serpins in arthropod biology[J].Semin Cell Dev Biol,2017,62:105-119.
[5]Law R H,Zhang Q,McGowan S,et al.An overview of the serpin superfamily[J].Genome Biol,2006,7(5):216.
[6]Irving J A,Pike R N,Lesk A M,et al.Phylogeny of the serpin superfamily:implications of patterns of amino acid conservation for structure and function[J].Genome Res,2000,10(12):1845-1864.
[7]Gulley M M,Zhang X,Michel K.The roles of serpins in mosquito immunology and physiology[J].J Insect Physiol,2013,59(2):138-147.
[8]PáleníkováJ,LieskovskáJ,LanghansováH,et al.Ixodes ricinus salivary serpin IRS-2 affects Th17differentiation via inhibition of the interleukin-6/STAT-3 signaling pathway[J].Infect Immun,2015,83(5):1949-1956.
[9]赵丽芳,陶美林,潘国庆.丝氨酸蛋白酶抑制剂超家族的研究进展[J].蚕业科学,2016,42(03):532-540.
[10]Rau J C,Beaulieu L M,Huntington J A,et al.Serpins in thrombosis,hemostasis and fibrinolysis[J].J Thromb Haemost.2007,5(suppl1):102-115.
[11]Francischetti I M,Sa-Nunes A,Mans B J,et al.The role of saliva in tick feeding[J].Front Biosci(Landmark Ed),2009,14:2051-2088.
[12]Leboulle G,Crippa M,Decrem Y,et al.Characterization of a novel salivary immunosuppressive protein from Ixodes ricinus ticks[J].J Biol Chem,2002,277(12):10083-10089.
[13]Prevot P P,Adam B,Boudjeltia K Z,et al.Antihemostatic effects of a serpin from the saliva of the tick Ixodes ricinus[J].J Biol Chem,2006,281(36):26361-26369.
[14]Anderson J M,Sonenshine D E,Valenzuela J G.Exploring the mialome of ticks:an annotated catalogue of midgut transcripts from the hard tick,Dermacentor variabilis(Acari:Ixodidae)[J].BMC Genomics,2008,9:552.
[15]Pak S C,Kumar V,Tsu C,et al.SRP-2 is a cross-class inhibitor that participates in postembryonic development of the nematode Caenorhabditis elegans:initial characterization of the clade L serpins[J].J Biol Chem,2004,279(15):15448-15459.
[16]Rodriguez-Valle M,Vance M,Moolhuijzen P M,et al.Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3(RMS-3)[J].Ticks Tick Borne Dis,2012,3(3):159-169.
[17]邓振,何庆丰,彭礼飞.中性粒细胞丝氨酸蛋白酶及其抑制剂与炎症[J].中国药理学通报,2014,30(3):314-317.
[18]Bode W.The structure of thrombin:a janus-headed proteinase[J].Semin Thromb Hemost,2006,32(Suppl 1):16-31.
[19]Prevot P P,Couvreur B,Denis V,et al.Protective immunity against Ixodes ricinus induced by a salivary serpin[J].Vaccine,2007,25(17):3284-3292.
[20]Prevot P P,Beschin A,Lins L,et al.Exosites mediate the anti-inflammatory effects of a multifunctional serpin from the saliva of the tick Ixodes ricinus[J].FEBS J,2009,276(12):3235-3246.
[21]Abraham E G,Pinto S B,Ghosh A,et al.An immuneresponsive serpin,SRPN6,mediates mosquito defense against malaria parasites[J].Proc Natl Acad Sci USA,2005,102(45):16327-16332.
[22]Pinto S B,Kafatos F C,Michel K.The parasite invasion marker SRPN6 reduces sporozoite numbers in salivary glands of Anopheles gambiae[J].Cell Microbiol,2008,10(4):891-898.
[23]Luo L M,Zhao L,Wen H L,et al.Haemaphysalis longicornis ticks as reservoir and vector of severe fever with Thrombocytopenia Syndrome Virus in China[J].Emerg Infect Dis,2015,21(10):1770-1776.
[24]Yu Y,Cao J,Zhou Y,et al.Isolation and characterization of two novel serpins from the tick Rhipicephalus haemaphysaloides[J].Ticks Tick Borne Dis,2013,4(4):297-303.