Advertisement

The kallikrein-like activity of Heloderma venom is inhibited by carbon monoxide

  • Vance G. NielsenEmail author
  • Nathaniel Frank
Article

Abstract

Lizards in the genus Heloderma are the most ancient venomous reptiles, with a traceable lineage nearly 100 million years old. The proteome of the venom of three of the remaining species (Heloderma suspectum, H. exasperatum, H. horridum) are very conserved, with kallikrein-like activity present to cause critical hypotension to immobilize and outright kill prey. Kallikrein-like activity would be expected to activate the contact protein pathway of coagulation, which would be detectable with thrombelastography in human plasma. Thus, it was proposed to determine if kallikrein-like activity could be detected with thrombelastography, and if this activity could be inhibited by carbon monoxide (CO) via a putative heme-based mechanism. Procoagulant activity of each venom was assessed via thrombelastography with normal plasma, and kallikrein-like activity confirmed with FX-depleted plasma. Venom was then exposed to carbon monoxide releasing molecule-2 (CORM-2) or its inactive releasing molecule to assess CO inhibition. All three venoms demonstrated kallikrein-like activity with the same potency and inhibition of activity by CO. In conclusion, the present work documented that procoagulant, kallikrein-like activity containing venoms of the oldest species of venomous reptiles was inhibited by CO, potentially via heme modulation. This is also the first identification and characterization of a kallikrein-like enzyme utilizing coagulation factor-depleted plasma to assess venom that inflicts hypotension. Future investigations will continue to define the vulnerability of venom enzymatic activities to CO.

Keywords

Kallikrein-like activity Phospholipase A2 Carbon monoxide Heme Thrombelastography 

Notes

Acknowledgements

We appreciate the technical assistance of Sarah A. Nielsen in the conduct of the thrombelastographic assays.

Funding

This investigation was supported by the Department of Anesthesiology, College of Medicine, at the University of Arizona.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This was an in vitro investigation and did not involve any living subjects.

References

  1. 1.
    Beaman KR, Beck DD, McGurty BM (2006) The beaded lizard (Heloderma horridum) and gila monster (Heloderma suspectum): a bibliography of the family Helodermatidae. Smithson Herpetol Inf Serv 136:1–66Google Scholar
  2. 2.
    Fry BG, Vidal N, Norman JA, Vonk FJ, Scheib H, Ramjan SF, Kuruppu S, Fung K, Hedges SB, Richardson MK, Hodgson WC, Ignjatovic V (2006) Early evolution of the venom system in lizards and snakes. Nature 439:584–588CrossRefGoogle Scholar
  3. 3.
    Douglas ME, Douglas MR, Schuett GW, Beck DD, Sullivan BK (2010) Conservation phylogenetics of helodermatid lizards using multiple molecular markers and a supertree approach. Mol Phylogenet Evol 55:153–167CrossRefGoogle Scholar
  4. 4.
    Reiserer RS, Schuett GW, Beck DD (2013) Taxonomic reassessment and conservation status of the beaded lizard, Heloderma horridum (Squamata: Helodermatidae). Amphib Reptile Conserv 7:74–96Google Scholar
  5. 5.
    Koludarov I, Jackson TN, Sunagar K, Nouwens A, Hendrikx I, Fry BG (2014) Fossilized venom: the unusually conserved venom profiles of Heloderma species (beaded lizards and gila monsters). Toxins 6:3582–3595CrossRefGoogle Scholar
  6. 6.
    Sanggaard KW, Dyrlund TF, Thomsen LR, Nielsen TA, Brøndum L, Wang T, Thøgersen IB, Enghild JJ (2015) Characterization of the gila monster (Heloderma suspectum suspectum) venom proteome. J Proteom 117:1–11CrossRefGoogle Scholar
  7. 7.
    Alagon A, Possani LD, Smart J, Schleuning WD (1986) Helodermatine, a kallikrein-like, hypotensive enzyme from the venom of Heloderma horridum horridum (Mexican beaded lizard). J Exp Med 164:1835–1845CrossRefGoogle Scholar
  8. 8.
    Sosa BP, Alagón AC, Martin BM, Possani LD (1986) Biochemical characterization of the phospholipase A2 purified from the venom of the Mexican beaded lizard (Heloderma horridum horridum Wiegmann). Biochemistry 25:2927–2933CrossRefGoogle Scholar
  9. 9.
    Datta G, Tu AT (1997) Structure and other chemical characterizations of gila toxin, a lethal toxin from lizard venom. J Pept Res 50:443–450CrossRefGoogle Scholar
  10. 10.
    Ivanov I, Matafonov A, Sun MF, Cheng Q, Dickeson SK, Verhamme M, Emsley J, Gailani D (2017) Proteolytic properties of single-chain factor XII: a mechanism for triggering contact activation. Blood 129:1527–1537CrossRefGoogle Scholar
  11. 11.
    Mohammed BM, Matafonov A, Ivanov I, Sun MF, Cheng Q, Dickeson SK, Li C, Sun D, Verhamme IM, Emsley J, Gailani D (2018) An update on factor XI structure and function. Thromb Res 161:94–105CrossRefGoogle Scholar
  12. 12.
    Nielsen VG, Cohen BM, Cohen E (2005) Effects of coagulation factor deficiency on plasma coagulation kinetics determined via thrombelastography: critical roles of fibrinogen and factors II, VII, X and XII. Acta Anaesthesiol Scand 49:222–231CrossRefGoogle Scholar
  13. 13.
    Nielsen VG (2019) Carbon monoxide inhibits the anticoagulant activity of phospholipase A2 purified from Crotalus adamanteus venom. J Thromb Thrombolysis 47:73–79CrossRefGoogle Scholar
  14. 14.
    Nielsen VG, Cerruti MA, Valencia OM, Amos Q (2016) Decreased snake venom metalloproteinase effects via inhibition of enzyme and modification of fibrinogen. Biometals 29:913–919CrossRefGoogle Scholar
  15. 15.
    Nielsen VG, Frank N (2019) Role of heme modulation in inhibition of Atheris, Atractaspis, Causus, Cerastes, Echis, and Macrovipera hemotoxic venom activity. Hum Exp Toxicol 38:216–226CrossRefGoogle Scholar
  16. 16.
    Nielsen VG, Frank N, Matika RW (2018) Carbon monoxide inhibits hemotoxic activity of Elapidae venoms: potential role of heme. Biometals 31:51–59CrossRefGoogle Scholar
  17. 17.
    Nielsen VG, Bazzell CM (2016) Carbon monoxide attenuates the effects of snake venoms containing metalloproteinases with fibrinogenase or thrombin-like activity on plasmatic coagulation. MedChemComm 7:1973–1979CrossRefGoogle Scholar
  18. 18.
    Nielsen VG, Bazzell CM (2017) Carbon monoxide releasing molecule-2 inhibition of snake venom thrombin-like activity: novel biochemical “brake”? J Thromb Thrombolysis 43:203–208CrossRefGoogle Scholar
  19. 19.
    Nielsen VG, Frank N (2018) Differential heme-mediated modulation of Deinagkistrodon, Dispholidus, Protobothrops and Pseudonaja hemotoxic venom activity in human plasma. Biometals 31:951–959CrossRefGoogle Scholar
  20. 20.
    Nielsen VG, Frank N, Matika RW (2018) Effects of heme modulation on Ovophis and Trimeresurus venom activity in human plasma. Toxins (Basel) 10:E322CrossRefGoogle Scholar
  21. 21.
    Nielsen VG, Losada PA (2017) Direct inhibitory effects of carbon monoxide on six venoms containing fibrinogenolytic metalloproteinases. Basic Clin Pharmacol Toxicol 120:207–212CrossRefGoogle Scholar
  22. 22.
    Suntravat M, Langlais PR, Sánchez EE, Nielsen VG (2018) CatroxMP-II: a heme-modulated fibrinogenolytic metalloproteinase isolated from Crotalus atrox venom. Biometals 31:585–593CrossRefGoogle Scholar
  23. 23.
    Nielsen VG (2018) Crotalus atrox venom exposed to carbon monoxide has decreased fibrinogenolytic activity in vivo in rabbits. Basic Clin Pharmacol Toxicol 122:82–86CrossRefGoogle Scholar
  24. 24.
    Murakami M, Taketomi Y, Sato H, Yamamoto K (2011) Secreted phospholipase A2 revisited. J Biochem 150:233–255CrossRefGoogle Scholar
  25. 25.
    Koludarov I, Jackson TN, Brouw BOD, Dobson J, Dashevsky D, Arbuckle K, Clemente CJ, Stockdale EJ, Cochran C, Debono J, Stephens C, Panagides N, Li B, Manchadi MR, Violette A, Fourmy R, Hendrikx I, Nouwens A, Clements J, Martelli P, Kwok HF, Fry BG (2017) Enter the dragon: the dynamic and multifunctional evolution of Anguimorpha lizard venoms. Toxins (Basel) 9:E242CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of AnesthesiologyThe University of Arizona College of MedicineTucsonUSA
  2. 2.MtoxinsOshkoshUSA

Personalised recommendations