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Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis

Ruo-Mei Cheng Xiao-Peng Tang Ai-Lin Long James Mwangi Ren Lai Rui-Pu Sun Cheng-Bo Long Zhen-Qing Zhang

Ruo-Mei Cheng, Xiao-Peng Tang, Ai-Lin Long, James Mwangi, Ren Lai, Rui-Pu Sun, Cheng-Bo Long, Zhen-Qing Zhang. Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis. Zoological Research, 2019, 40(3): 205-210. doi: 10.24272/j.issn.2095-8137.2019.037
Citation: Ruo-Mei Cheng, Xiao-Peng Tang, Ai-Lin Long, James Mwangi, Ren Lai, Rui-Pu Sun, Cheng-Bo Long, Zhen-Qing Zhang. Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis. Zoological Research, 2019, 40(3): 205-210. doi: 10.24272/j.issn.2095-8137.2019.037

菲牛蛭(Hirudinaria manillensis)中一种新型抗凝血多肽的分离纯化与表征

doi: 10.24272/j.issn.2095-8137.2019.037

Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis

Funds: This work was supported by the National Natural Science Foundation of China (21761142002, 81770464), Ministry of Science and Technology of China (2018ZX09301043-003), Chinese Academy of Sciences (QYZDJ-SSW-SMC012, SAJC201606), West Light Foundation and Youth Innovation Promotion Association (2017432), and Yunnan Provincial Science and Technology Department (2017FB038, 2015BC005)
More Information
    Corresponding author: Cheng-Bo Long,Zhen-Qing Zhang
  • 摘要: 目前关于菲牛蛭(Hirudinaria manillensis)中的蛋白酶抑制剂的报道还比较少。本研究纯化了一种新型抗凝血特性的蛋白酶抑制剂,命名为bdellin-HM-2;其分子量为1.4x104,分子内N端有三对二硫键和C端有多个HHXDD和HXDD基序。cDNA克隆结果揭示了bdellin-HM-2的可能的核苷酸编码蛋白由132个氨基酸组成,由 399 bp开放阅读框(ORF)编码。序列比对表明,bdellin-HM-2与“非典型”Kazal型丝氨酸蛋白酶抑制剂相似,但却对胰蛋白酶、弹性酶、糜蛋白酶、激肽酶、XIIa因子(FXIIa)、因子XIa (FXIa)、因子Xa (FXa)、凝血酶以及纤溶酶都没有抑制作用。Bdellin-HM-2是通过延长活化的部分凝血活酶时间(aPTT)达到抗凝血作用,继而获得从宿主那里吸食血液作为食物的能力。我们的研究结果表明,Bdellin-HM-2可能在这类水蛭的吸血过程中都起到重要作用,可能是一种临床上潜在的候选抗血栓药物。
  • [1]
    null Ali MF , Lips KR , Knoop FC , Fritzsch B , Miller C , Conlon JM . 2002. Antimicrobial peptides and protease inhibitors in the skin secretions of the crawfish frog, Rana areolata . Biochimica et Biophysica Acta-Proteins and Proteomics, 1601(1): 55–63.
    null Birrell GW , Earl STH , Wallis TP , Masci PP , De Jersey J , Gorman JJ , Lavin MF . 2007. The diversity of bioactive proteins in Australian snake venoms. Molecular & Cellular Proteomics, 6(6): 973–986.
    null Bode W , Huber R . 1992. Natural protein proteinase inhibitors and their interaction with proteinases. European Journal of Biochemistry, 204(2): 433–451.
    null De Marco R , Lovato DV , Torquato RJS , Clara RO , Buarque DS , Tanaka AS . 2010. The first pacifastin elastase inhibitor characterized from a blood sucking animal. Peptides, 31(7): 1280–1286.
    null Dodt J . 1995. Anticoagulatory substances of bloodsucking animals: from hirudin to hirudin mimetics. Angewandte Chemie-International Edition, 34(8): 867–880.
    null Eigenbrot C , Ultsch M , Lipari MT , Moran P , Lin SJ , Ganesan R , Quan C , Tom J , Sandoval W , Van Lookeren Campagne M , Kirchhofer D . 2012. Structural and functional analysis of HtrA1 and its subdomains. Structure, 20(6): 1040–1050.
    null Fink E , Rehm H , Gippner C , Bode W , Eulitz M , Machleidt W , Fritz H . 1986. The primary structure of Bdellin B-3 from the leech Hirudo Medicinalis - Bdellin B-3 is a compact proteinase inhibitor of a “Non-classical” Kazal Type - It is present in the leech in a high molecular mass form. Biological Chemistry Hoppe-Seyler, 367(12): 1235–1242.
    null Gupta R , Brunak S . 2002. Prediction of glycosylation across the human proteome and the correlation to protein function. Pacific Symposium on Biocomputing, 7(3): 310–322.
    null Hong SJ , Kang KW . 1999. Purification of granulin-like polypeptide from the blood-sucking leech, Hirudo nipponia . Protein Expression and Purification, 16(2): 340–346.
    null Kim YH , Choi JG , Lee GM , Kang KW . 2001. Domain and genomic sequence analysis of bdellin-KL, a leech-derived trypsin-plasmin inhibitor. Journal of Biochemistry, 130(3): 431–438.
    null Kim JY , Park SC , Hwang I , Cheong H , Nah JW , Hahm KS , Park Y . 2009. Protease inhibitors from plants with antimicrobial activity. International Journal of Molecular Sciences, 10(6): 2860–2872.
    null Lai Y , Li B , Liu W , Wang G , Du C , Ombati R , Lai R , Long C , Li H . 2016. Purification and characterization of a novel Kazal-type trypsin inhibitor from the leech of Hirudinaria manillensis . Toxins, 8(8): 229–239.
    null Laskowski M , Jr, Kato I . 1980. Protein inhibitors of proteinases. Annual Review of Biochemistry, 49: 593–626.
    null Leung D , Abbenante G , Fairlie DP . 2000. Protease inhibitors: current status and future prospects. Journal of Medicinal Chemistry, 43(3): 305–341.
    null M?gert HJ , St?ndker L , Kreutzmann P , Zucht HD , Reinecke M , Sommerhoff CP , Fritz H , Forssmann WG . 1999. LEKTI, a novel 15-domain type of human serine proteinase inhibitor. Journal of Biological Chemistry, 274(31): 21499–21502.
    null Markwardt F . 1996. State-of-the-Art Review: Antithrombotic agents from hematophagous animals. Clinical and Applied Thrombosis-Hemostasis, 2(2): 75–82.
    null Markwardt F . 2002. Hirudin as alternative anticoagulant-a historical review. Seminarsin Thrombosis and Hemostasis, 28(5): 405–413.
    null Mende K , Petoukhova O , Koulitchkova V , Schaub GA , Lange U , Kaufmann R , Nowak G . 1999. Dipetalogastin, a potent thrombin inhibitor from the blood-sucking insect Dipetalogaster maximus-cDNA cloning, expression and characterization. European Journal of Biochemistry, 266(2): 583–590.
    null Moser M , Auerswald E , Mentele R , Eckerskorn C , Fritz H , Fink E . 1998. Bdellastasin, a serine protease inhibitor of the antistasin family from the medical leech (Hirudo medicinalis) - primary structure, expression in yeast, and characterisation of native and recombinant inhibitor. European Journal of Biochemistry, 253(1): 212–220.
    null Ohtsubo K , Marth JD . 2006. Glycosylation in cellular mechanisms of health and disease. Cell, 126(5): 855–867.
    null Rimphanitchayakit V , Tassanakajon A . 2010. Structure and function of invertebrate Kazal-type serine proteinase inhibitors. Developmental & Comparative Immunology, 34(4): 377–386.
    null Ryan CA . 1990. Protease inhibitors in plants-genes for improving defenses against insects and pathogens. Annual Review of Phytopathology, 28: 425–449.
    null Sabotic J , Kos J . 2012. Microbial and fungal protease inhibitors-current and potential applications. Applied Microbiology and Biotechnology, 93(4): 1351–1375.
    null Salzet M . 2001. Anticoagulants and inhibitors of platelet aggregation derived from leeches. FEBS Letters, 492(3): 187–192.
    null Salzet M , Chopin V , Baert JL , Matias I , Malecha J . 2000. Theromin, a novel leech thrombin inhibitor. Journal of Biological Chemistry, 275(40): 30774–30780.
    null Shadrin N , Shapira MG , Khalfin B , Uppalapati L , Parola AH , Nathan I . 2015. Serine protease inhibitors interact with IFN-γ through up-regulation of FasR; a novel therapeutic strategy against cancer. Experimental Cell Research, 330(2): 233–239.
    null Strube KH , Kr?ger B , Bialojan S , Otte M , Dodt J . 1993. Isolation, sequence-analysis, and cloning of Haemadin - an anticoagulant peptide from the Indian leech. Journal of Biological Chemistry, 268(12): 8590–8595.
    null Supuran CT , Scozzafava A , Clare BW . 2002. Bacterial protease inhibitors. Medicinal Research Reviews, 22(4): 329–372.
    null Tuszynski GP , Gasic TB , Gasic GJ . 1987. Isolation and characterization of antistasin -an inhibitor of metastasis and coagulation. Journal of Biological Chemistry, 262(20): 9718–9723.
    null Van De Locht A , Lamba D , Bauer M , Huber R , Friedrich T , Kr?ger B , H?ffken W , Bode W . 1995. Two heads are better than one: crystal structure of the insect derived double domain Kazal inhibitor rhodniin in complex with thrombin. EMBO Journal, 14(21): 5149–5157.
    null Vicu?a L , Strochlic DE , Latremoliere A , Bali KK , Simonetti M , Husainie D , Prokosch S , Riva P , Griffin RS , Njoo C , Gehrig S , Mall MA , Arnold B , Devor M , Woolf CJ , Liberles SD , Costigan M , Kuner R . 2015. The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell-derived leukocyte elastase. Nature Medicine, 21(5): 518–523.
    null Wang LJ , Gao GF , Bai SP , Liu ZG , Liang JG , Zhang KY , Lai R . 2005. A thrombin inhibitor from ixodid tick, Ixodide sinesis . Zoological Research, 26(3): 328–331.
    null Zhang Y . 2006. Amphibian skin secretions and bio-adaptive significance- implications from Bombina maxima skin secretion proteome. Zoological Research, 27(1): 101–112.
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  • 收稿日期:  2018-09-03
  • 刊出日期:  2019-05-18

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