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程若梅, 唐小芃, 龙爱琳, Mwangi James, 赖仞, 孙瑞璞, 龙承波, 张真庆. 2019: 菲牛蛭(Hirudinaria manillensis)中一种新型抗凝血多肽的分离纯化与表征. 动物学研究, 40(3): 205-210. DOI: 10.24272/j.issn.2095-8137.2019.037
引用本文: 程若梅, 唐小芃, 龙爱琳, Mwangi James, 赖仞, 孙瑞璞, 龙承波, 张真庆. 2019: 菲牛蛭(Hirudinaria manillensis)中一种新型抗凝血多肽的分离纯化与表征. 动物学研究, 40(3): 205-210. DOI: 10.24272/j.issn.2095-8137.2019.037
Ruo-Mei Cheng, Xiao-Peng Tang, Ai-Lin Long, James Mwangi, Ren Lai, Rui-Pu Sun, Cheng-Bo Long, Zhen-Qing Zhang. 2019: Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis. Zoological Research, 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. 2019: Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis. Zoological Research, 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

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

  • 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。

     

    Abstract: Protease inhibitors have been reported rarely from the leech Hirudinaria manillensis. In this study, we purified a novel protease inhibitor (bdellin-HM-2) with anticoagulant properties from H. manillensis. With a molecular weight of 1.4x104, bdellin-HM-2 was also characterized with three intra-molecular disulfide bridges at the N-terminus and multiple HHXDD and HXDD motifs at the C-terminus. cDNA cloning revealed that the putative nucleotide-encoding protein of bdellin-HM-2 contained 132 amino acids and was encoded by a 399 bp open reading frame (ORF). Sequence alignment showed that bdellin-HM-2 shared similarity with the “non-classical” Kazal-type serine protease inhibitors, but had no inhibitory effect on trypsin, elastase, chymotrypsin, kallikrein, factor XIIa (FXIIa), factor XIa (FXIa), factor Xa (FXa), thrombin, or plasmin. Bdellin-HM-2 showed anticoagulant effects by prolonging the activated partial thromboplastin time (aPTT), indicating a role in enabling H. manillensis to obtain a blood meal from its host. Our results suggest that bdellin-HM-2 may play a crucial role in blood-sucking in this leech species and may be a potential candidate for the development of clinical anti-thrombotic drugs.

     

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