title: Acetylene inhibition of Azotobacter vinelandii hydrogenase: acetylene binds tightly to the large subunit.
abstract: Acetylene is a slow-binding inhibitor of the Ni- and Fe-containing dimeric hydrogenase isolated from Azotobacter vinelandii. Acetylene was released from hydrogenase during the recovery from inhibition. This indicates that no transformation of acetylene to another compound occurred as a result of the interaction with hydrogenase. However, the release of C2H2 proceeds more rapidly than the recovery of activity, which indicates that release of C2H2 is not sufficient for recovery of activity. Acetylene binds tightly to native hydrogenase; hydrogenase and radioactivity coelute from a gel permeation column following inhibition with 14C2H2. Acetylene, or a derivative, remains bound to the large 65,000 MW subunit (and not to the small 35,000 MW subunit) of hydrogenase following denaturation as evidenced by SDS-PAGE and fluorography of 14C2H2-inhibited hydrogenase. This result suggests that C2H2, and by analogy H2, binds to and is activated by the large subunit of this dimeric hydrogenase. Radioactivity is lost from 14C2H2-inhibited protein during recovery. The inhibition is remarkably specific for C2H2: propyne, butyne, and ethylene are not inhibitors.
authors.affiliation: Laboratory for Nitrogen Fixation Research, Oregon State University, Corvallis 97331-2902.
authors.forename: J H
authors.initials: JH
authors.lastname: Sun
authors.forename: M R
authors.initials: MR
authors.lastname: Hyman
authors.forename: D J
authors.initials: DJ
authors.lastname: Arp
mesh_terms: D000114:Acetylene / Q000737:chemistry* / Q000494:pharmacology; D016948:Azotobacter vinelandii / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology*; D001667:Binding, Competitive; D002250:Carbon Radioisotopes; D006864:Hydrogenase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification; D007700:Kinetics; D011485:Protein Binding; D011487:Protein Conformation; D013379:Substrate Specificity
chemical_list: D002250:Carbon Radioisotopes; D006864:Hydrogenase; D000114:Acetylene