Joannsin, a novel Kunitz-type FXa inhibitor from the venom of Prospirobolus joannsi
09 November 2016
Accepted after major revision: 19 February 2017
07 November 2017 (online)
The repugnatorial glands of millipedes release various defensive chemical secretions. Although varieties of such defensive secretions have been studied, none of them is protein or peptide. Herein, a novel factor Xa (FXa) inhibitor named joannsin was identified and characterised from repugnatorial glands of Prospirobolus joannsi. Joannsin is composed of 72 amino acid residues including six cysteines, which form three intra-molecular disulfide bridges. It is a member of Kunitz-type protease inhibitor family, members of which are also found in the secretory glands of other arthropods. Recombinant joannsin exhibited remarkable inhibitory activity against trypsin and FXa with a Ki of 182.7 ± 14.6 and 29.5 ± 4.7 nM, respectively. Joannsin showed strong anti-thrombosis functions in vitro and in vivo. Joannsin is the first peptide component in millipede repugnatorial glands to be identified and is a potential candidate and/or template for the development of anti-thrombotic agents. These results also indicated that there is Kunitz-type protease inhibitor toxin in millipede repugnatorial glands as in other arthropods secretory glands.
§These authors contributed equally to this paper.
- 1 Zhang Y. Why do we study animal toxins?. Zool Res 2015; 36: 183-222.
- 2 Abdul HM, Yang S, Ren L. Centipede Venoms and Their Components: Resources for Potential Therapeutic Applications. Toxins 2015; 7: 4832-4851.
- 3 Eisner T, Meinwald J, Attyqalle AB. et al. Rendering the inedible edible: circumvention of a millipede’s chemical defense by a predaceous beetle larva. Proc Natl Acad Sci USA 1998; 95: 1108-1113.
- 4 Dadashipour M, Ishida Y, Yamamoto K. et al. Discovery and molecular and bio-catalytic properties of hydroxynitrile lyase from an invasive millipede, Chamber-linius hualienensis . Proc Natl Acad Sci USA 2015; 112: 10605-10610.
- 5 Kuwahara Y, Ichiki Y, Morita M. et al. Chemical polymorphism in defense secretions during ontogenetic development of the millipede Niponia nodulosa . J Chem Ecol 2015; 41: 15-21.
- 6 Gubb D, Sanz-Parra A, Barcena L. et al. Protease inhibitors and proteolytic signalling cascades in insects. Biochimie 2010; 92: 1749-1759.
- 7 Peng K, Yi K, Lei Z. et al. Two novel antimicrobial peptides from centipede venoms. Toxicon 2010; 55: 274-279.
- 8 Yang S, Liu Z, Xiao Y. et al. Chemical punch packed in venoms makes centipedes excellent predators. Mol Cell Proteomics 2012; 11: 640-650.
- 9 Ma H, Xiao-Peng T, Yang SL. et al. Protease inhibitor in scorpion (Mesobuthus eupeus) venom prolongs the biological activities of the crude venom. Chin J Nat Med 2016; 14: 607-614.
- 10 Gulliani GL, Hyun BH, Litten MB. Blood recalcification time. A simple and reliable test to monitor heparin therapy. Am J Clin Pathol 1976; 65: 390-396.
- 11 He W, Wu JJ, Ning J. et al. Inhibition of human cytochrome P450 enzymes by licochalcone A, a naturally occurring constituent of licorice. Toxicol In Vitro 2015; 29: 1569-1576.
- 12 Jung WK, Kim SK. Isolation and characterisation of an anticoagulant oligopep-tide from blue mussel, Mytilus edulis . Food Chem 2009; 117: 687-692.
- 13 Jing T, Fang Y, Han Y. et al. YY-39, a tick anti-thrombosis peptide containing RGD domain. Peptides 2015; 68: 99-104.
- 14 Sasaki SD, Cotrin SS, Carmona AK. et al. An unexpected inhibitory activity of Kunitz-type serine proteinase inhibitor derived from Boophilus microplus trypsin inhibitor on cathepsin L. Biochem Biophys Res Commun 2006; 341: 266-272.
- 15 Liao M, Zhou J, Gong H. et al. Hemalin, a thrombin inhibitor isolated from a midgut cDNA library from the hard tick Haemaphysalis longicornis . J Insect Physiol 2009; 55: 164-173.
- 16 Macedo-Ribeiro S, Almeida C, Calisto BM. et al. Isolation, cloning and structural characterisation of boophilin, a multifunctional Kunitz-type proteinase inhibitor from the cattle tick. PLoS One 2007; 3: e1624.
- 17 Ren L, Takeuchi H, Jonczy J. et al. A thrombin inhibitor from the ixodid tick, Amblyomma hebraeum . Gene 2004; 342: 243-249.
- 18 von Reumont BM, Blanke A, Richter S. et al. The first venomous crustacean revealed by transcriptomics and functional morphology: remipede venom glands express a unique toxin cocktail dominated by enzymes and a neurotoxin. Mol Biol Evol 2014; 31: 48-58.
- 19 Ma D, Wang Y, Yang H. et al. Anti-thrombosis repertoire of blood-feeding horsefly salivary glands. Mol Cell Proteomics 2009; 8: 2071-2079.
- 20 Xu X, Yang H, Ma D. et al. Toward an understanding of the molecular mechanism for successful blood feeding by coupling proteomics analysis with pharmacological testing of horsefly salivary glands. Mol Cell Proteomics 2008; 7: 582-590.
- 21 Rong M, Liu J, Zhang M. et al. A sodium channel inhibitor ISTX-I with a novel structure provides a new hint at the evolutionary link between two toxin folds. Sci Rep 2016; 6: 29691.
- 22 Ribeiro JM, Makoul GT, Levine J. et al. Antihemostatic, antiinflammatory, and immunosuppressive properties of the saliva of a tick, Ixodes dammini . J Exp Med 1985; 161: 332-344.
- 23 Zhou Z, Yang H, Xu X. et al. The first report of kininogen from invertebrates. Biochem Biophys Res Commun 2006; 347: 1099-1102.
- 24 Yang H, Xu X, Ma D. et al. A phospholipase A1 platelet activator from the wasp venom of Vespa magnifica (Smith). Toxicon 2008; 51: 289-296.
- 25 Smith JJ, Hill JM, Little MJ. et al. Unique scorpion toxin with a putative ancestral fold provides insight into evolution of the inhibitor cystine knot motif. Proc Natl Acad Sci USA 2011; 108: 10478-10483.
- 26 Wang XH, Connor M, Smith R. et al. Discovery and characterization of a family of insecticidal neurotoxins with a rare vicinal disulfide bridge. Nat Struct Biol 2000; 7: 505-513.
- 27 Yang S, Yang F, Wei N. et al. A pain-inducing centipede toxin targets the heat activation machinery of nociceptor TRPV1. Nat Commun 2015; 6: 284a-284a.
- 28 Yang S, Xiao Y, Kang D. et al. Discovery of a selective NaV1.7 inhibitor from centipede venom with analgesic efficacy exceeding morphine in rodent pain models. Proc Natl Acad Sci USA 2013; 110: 17534-17539.
- 29 Cao Z, Yu Y, Wu Y. et al. The genome of Mesobuthus martensii reveals a unique adaptation model of arthropods. Nat Commun. 2011; 4: 2602-2602.
- 30 Smith JJ, Hill JM, Little MJ. et al. Unique scorpion toxin with a putative ancestral fold provides insight into evolution of the inhibitor cystine knot motif. Proc Natl Acad Sci USA 2011; 108: 10478-10483.
- 31 Wang XH, Connor M, Smith R. et al. Discovery and characterization of a family of insecticidal neurotoxins with a rare vicinal disulfide bridge. Nat Struct Biol 2000; 7: 505-513.
- 32 Liu Z, Cai T, Zhu Q. et al. Structure and function of hainantoxin-III, a selective antagonist of neuronal tetrodotoxin-sensitive voltage-gated sodium channels isolated from the Chinese bird spider Ornithoctonus hainana . J Biol Chem 2013; 288: 20392-20403.
- 33 Mourão CB, Schwartz EF. Protease inhibitors from marine venomous animals and their counterparts in terrestrial venomous animals. Mar Drugs 2013; 11: 2069-2112.
- 34 Siemens J, Zhou S, Piskorowski R. et al. Spider toxins activate the capsaicin receptor to produce inflammatory pain. Nature 2006; 444: 208-212.
- 35 Bohlen CJ, Priel A, Zhou S. et al. A bivalent tarantula toxin activates the capsaicin receptor, TRPV1, by targeting the outer pore domain. Cell. 2010; 141: 834-845.
- 36 Branco VG, Iqbal A, Alvarez-Flores MP. et al. Amblyomin-X having a Kunitz-type homologous domain, is a noncompetitive inhibitor of FXa and induces anticoagulation in vitro and in vivo. Biochim Biophys Acta 2016; 1864: 1428-1435.
- 37 Huang ZF, Wun TC, Broze Jr GJ. Kinetics of factor Xa inhibition by tissue factor pathway inhibitor. J Biol Chem 1993; 268: 26950-26955.