Purification and characterization of inorganic pyrophosphatase from Bacillus stearothermophilus.
Inorganic pyrophosphatase [EC 3.6.1.1] was purified from Bacillus stearothermophilus to a homogeneous state both ultracentrifugally and electrophoretically. Ultracentrifugal analysis revealed that the molecular weight of the enzyme is 122,000 and the sedimentation coefficient (S0.34%/20, W) is 5.2S. The enzyme molecule in 0.1% sodium dodecylsulfate solution containing 1 mM 2-mercaptoethanol had an estimated molecular weight of 70,000 on the basis of SDS-polyacrylamide gel electrophoresis results, which indicates that the enzyme may consist of two subunits. Divalent cations such as Mg2+, Mn2+, and Co2+ are required for the enzymatic activity. Pyrophosphate is the only substrate for the enzyme. ATP and p-chloromercuribenzoate inhibit the enzyme reaction markedly.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D011485:Protein Binding; D011755:Pyrophosphatases / Q000302:isolation & purification / Q000378:metabolism*
D002729:Chloromercuribenzoates; D046911:Macromolecular Substances; D003035:Cobalt; D008345:Manganese; D000255:Adenosine Triphosphate; D011755:Pyrophosphatases; D008274:Magnesium