An unusual thiol-driven fumarate reductase in Methanobacterium with the production of the heterodisulfide of coenzyme M and N-(7-mercaptoheptanoyl)threonine-O3-phosphate.
An unusual fumarate reductase was purified from cell extracts of Methanobacterium thermoautotrophicum and partially characterized. Two coenzymes previously isolated from cell extracts, 2-mercaptoethane-sulfonic acid (HS-CoM) and N-(7-mercaptoheptanoyl)threonine-O3-phosphate (HS-HTP), were established as direct electron donors for fumarate reductase. By measuring the consumption of free thiol, we determined that fumarate reductase catalyzed the oxidation of HS-CoM and HS-HTP; by the direct measurement of succinate and the heterodisulfide of HS-CoM and HS-HTP (CoM-S-S-HTP), we established that these compounds were products of the fumarate reductase reaction. A number of thiol-containing compounds did not function as substrates for fumarate reductase, but this enzyme had high specific activity when HS-CoM and HS-HTP were used as electron donors. HS-CoM and HS-HTP were quantitatively oxidized by the fumarate reductase reaction, and results indicated that this reaction was irreversible. Additionally, by measuring formylmethanofuran, we demonstrated that the addition of fumarate to cell extracts activated CO2 fixation for the formation of formylmethanofuran. Results indicated that this activation resulted from the production of CoM-S-S-HTP (a compound known to be involved in the activation of formylmethanofuran synthesis) by the fumarate reductase reaction.
Bobik
T A
TA
Department of Microbiology, University of Illinois, Urbana 61801.
Wolfe
R S
RS
D004220:Disulfides / Q000378:metabolism; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D019605:Euryarchaeota / Q000201:enzymology*; D005650:Fumarates / Q000494:pharmacology; D005663:Furans / Q000378:metabolism; D008623:Mercaptoethanol; D015080:Mesna / Q000378:metabolism*; D010084:Oxidation-Reduction; D010769:Phosphothreonine / Q000378:metabolism / Q000031:analogs & derivatives; D013379:Substrate Specificity; D013385:Succinate Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D013386:Succinates / Q000378:metabolism; D019802:Succinic Acid; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D004220:Disulfides; D005650:Fumarates; D005663:Furans; D013386:Succinates; D013438:Sulfhydryl Compounds; C049349:7-mercaptoheptanoylthreonine phosphate; D010769:Phosphothreonine; D008623:Mercaptoethanol; C032005:fumaric acid; C050842:formylmethanofuran; D019802:Succinic Acid; D013385:Succinate Dehydrogenase; D015080:Mesna