Effect of molybdenum and tungsten on synthesis and composition of formate dehydrogenase in Methanobacterium formicicum.
The influence of sodium molybdate and sodium tungstate on formate dehydrogenase activity was studied in H2-CO2-grown cultures of Methanobacterium formicicum. Depletion of molybdate from the growth medium resulted in a 75-fold decrease of intracellular molybdenum and a 35-fold decrease in enzyme activity; however, growth rate and cell yields were not influenced. By using an indirect enzyme-linked immunoassay, the amount of formate dehydrogenase approximated 3% of the total protein in cells grown in the presence of molybdate. Molybdenum-starved cells contained approximately 15-fold less formate dehydrogenase protein; Western blot (immunoblot) analysis revealed that both subunits of the enzyme were synthesized. Molybdenum starvation resulted in an increase in the amount of mRNA that hybridized to fdh-specific DNA. The results indicated an inverse relationship between the amount of transcript and the amount of formate dehydrogenase protein detected in response to molybdenum starvation. The addition of 1 mM tungstate to molybdate-containing media resulted in nearly complete loss of enzyme activity and decreased the intracellular concentration of molybdenum 10-fold. Cells grown in the presence of tungstate synthesized high amounts of inactive formate dehydrogenase and contained mRNA that hybridized to fdh-specific DNA in amounts similar to that in cells grown with sufficient molybdate. Inactive formate dehydrogenase, purified from cells grown in the presence of tungstate, had the same subunit composition and contained amounts of molybdopterin cofactor, albeit metal-free, comparable to those in the active enzyme.
May
H D
HD
Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Patel
P S
PS
Ferry
J G
JG
D000445:Aldehyde Oxidoreductases / Q000096:biosynthesis*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D019605:Euryarchaeota / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D005560:Formate Dehydrogenases / Q000096:biosynthesis* / Q000235:genetics; D005561:Formates / Q000378:metabolism; D007118:Immunoassay; D008982:Molybdenum / Q000494:pharmacology*; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D014414:Tungsten / Q000494:pharmacology*
D005561:Formates; D012329:RNA, Bacterial; D012333:RNA, Messenger; C030544:formic acid; D008982:Molybdenum; D005560:Formate Dehydrogenases; D000445:Aldehyde Oxidoreductases; D014414:Tungsten