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

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 1.4x10(4), bdellin-HM-2 was also characterized with three intra-mol...

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Detalles Bibliográficos
Autores principales: Cheng, Ruo-Mei, Tang, Xiao-Peng, Long, Ai-Lin, Mwangi, James, Lai, Ren, Sun, Rui-Pu, Long, Cheng-Bo, Zhang, Zhen-Qing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Science Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6591161/
https://www.ncbi.nlm.nih.gov/pubmed/31011131
http://dx.doi.org/10.24272/j.issn.2095-8137.2019.037
Descripción
Sumario: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 1.4x10(4), 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.