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Hirudins of the Asian medicinal leech, Hirudinaria manillensis: same same, but different

  • Genetics, Evolution, and Phylogeny - Original Paper
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Abstract

Blood coagulation in vertebrates is a complex mechanism that involves the precisely coordinated and regulated action of a cascade of factors in order to prevent excessive blood loss upon wounding. Any blood sucking ectoparasite, however, has to circumvent this mechanism to ensure the uptake of an adequate blood meal. Inhibitors of blood coagulation in the saliva are hence widespread among these animals. Thrombin as a key factor of blood coagulation is a prominent target of such inhibitors, and hirudin is probably the best known among the thrombin inhibitors. Hirudin was originally described in the genus Hirudo, but occurs in other leech genera like Hirudinaria and Macrobdella as well. Besides several isoforms of hirudin, a new class of putative leech saliva components, the hirudin-like factors (HLFs), was identified in both genera Hirudo and Hirudinaria. Here, we describe the expression, purification, and functional characterization of three HLFs (HLF5, 6, and 8, respectively) and two additional hirudins (HM3 and HM4) of Hirudinaria manillensis. While HLF6 lacked any inhibitory activity on thrombin, HLF5 as well as HLF8 clearly exhibited anticoagulatory properties. The inhibitory activity of HLF5 and HLF8, however, was much lower compared with both HM3 and HM4 of Hirudinaria manillensis as well as the hirudin variants 1 (HV1) and 2 (HV2) of Hirudo medicinalis. Neither an inhibition of trypsin nor a platelet aggregation was caused by HLF8. Our data indicates the presence of two classes (rather than isoforms) of hirudins in Hirudinaria manillensis with markedly different inhibitory activity on human thrombin.

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Acknowledgments

The authors would like to thank Detlef Menzel (Futura-Egel-Zucht, Potsdam) for providing the Hirudo specimens and Bethany Sawyer (Biopharm U.K. Ltd., Swansea) for providing the Hirudinaria specimens. We are thankful to Sabine Ziesemer and Amanda Wiesenthal for critical reading of the manuscript and to the reviewers for their constructive and helpful comments.

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  1. Animal Physiology and Biochemistry, Zoological Institute and Museum, University of Greifswald, Felix-Hausdorff-Str. 1, 17489, Greifswald, Germany

    Phil Lukas, Jan-Peter Hildebrandt & Christian Müller

  2. Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Felix-Hausdorff-Str. 3, 17489, Greifswald, Germany

    Robert Wolf & Bernhard H. Rauch

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We declare that the experiments described in this paper comply with the current laws in Germany. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures in our studies involving human participants (platelet aggregation experiments) were approved by the local ethics committee and performed according to the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Lukas, P., Wolf, R., Rauch, B.H. et al. Hirudins of the Asian medicinal leech, Hirudinaria manillensis: same same, but different. Parasitol Res 118, 2223–2233 (2019). https://doi.org/10.1007/s00436-019-06365-z

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