Carbonic anhydrase metabolism is a key factor in the toxicity of CO2 and COS but not CS2 toward the flour beetle Tribolium castaneum [Coleoptera: Tenebrionidae].
The analogues carbon dioxide (CO(2)), carbonyl sulfide (COS) and carbon disulfide (CS(2)) have been useful as substrate probes for enzyme activities. Here we explored the affinity of the enzyme carbonic anhydrase for its natural substrate CO(2), as well as COS and CS(2) (1) by in vitro kinetic metabolism studies using pure enzyme and (2) through mortality bioassay of insects exposed to toxic levels of each of the gases during carbonic anhydrase inhibition. Hydrolysis of COS to form hydrogen sulfide was catalysed rapidly showing parameters K(m) 1.86 mM and K(cat) 41 s(-1) at 25 degrees C; however, the specificity constant (K(cat)/K(m)) was 4000-fold lower than the reported value for carbonic anhydrase-catalysed hydration of CO(2). Carbonic anhydrase-mediated CS(2) metabolism was a further 65,000-fold lower than COS. Both results demonstrate the deactivating effect toward the enzyme of sulfur substitution for oxygen in the molecule. We also investigated the role of carbonic anhydrases in CO(2), COS and CS(2) toxicity using a specific inhibitor, acetazolamide, administered to Tribolium castaneum (Herbst) larvae via the diet. CO(2) toxicity was greatly enhanced by up to seven-fold in acetazolamide-treated larvae indicating that carbonic anhydrases are a key protective enzyme in elevated CO(2) concentrations. Conversely, mortality was reduced by up to 12-fold in acetazolamide-treated larvae exposed to COS due to reduced formation of toxic hydrogen sulfide. CS(2) toxicity was unaffected by acetazolamide. These results show that carbonic anhydrase has a key role in toxicity of the substrates CO(2) and COS but not CS(2), despite minor differences in chemical formulae.
Haritos
Victoria S
VS
CSIRO Entomology, GPO Box 1700, Canberra, ACT 2602, Australia.
Dojchinov
Greg
G
D000818:Animals; D002245:Carbon Dioxide / Q000633:toxicity*; D002246:Carbon Disulfide / Q000633:toxicity*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D013461:Sulfur Oxides / Q000633:toxicity*; D014231:Tribolium / Q000187:drug effects* / Q000201:enzymology* / Q000254:growth & development
D013461:Sulfur Oxides; D002245:Carbon Dioxide; C010063:carbonyl sulfide; D002256:Carbonic Anhydrases; D002246:Carbon Disulfide