Volume 40 Issue 3
May  2019
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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

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

doi: 10.24272/j.issn.2095-8137.2019.037
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)
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  • Corresponding author: Cheng-Bo Long,Zhen-Qing Zhang
  • Received Date: 2018-09-03
  • Publish Date: 2019-05-18
  • 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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Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis

doi: 10.24272/j.issn.2095-8137.2019.037
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)
    Corresponding author: Cheng-Bo Long,Zhen-Qing Zhang

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.

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
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