S-adenosylmethionine decarboxylase from the thermophilic archaebacterium Sulfolobus solfataricus. Purification, molecular properties and studies on the covalently bound pyruvate.
S-Adenosylmethionine decarboxylase from Sulfolobus solfataricus, a thermoacidophilic archaebacterium optimally growing at 87 degrees C, has been purified to homogeneity. The specific activity of the homogeneous enzyme is 12 nmol CO2 formed min-1 (mg protein)-1 and the overall yield 8%. The enzyme is thermophilic with an optimum at 75 degrees C, is thermostable, and does not require divalent cations or putrescine for activity. It has a molecular mass of 32 kDa, and appears to be a monomeric protein. S-Adenosylmethionine decarboxylase from S. solfataricus contains covalently linked pyruvate as prosthetic group and is inactivated in a time-dependent process by NaCNBH3, in the presence of both the substrate and the product. Incubation with decarboxylated S-adenosyl[Me-3H]methionine and NaCNBH3 resulted in the labeling of the protein at the active site.
Cacciapuoti
G
G
Institute of Biochemistry of Macromolecules, First Medical School, University of Naples, Italy.
Porcelli
M
M
De Rosa
M
M
Gambacorta
A
A
Bertoldo
C
C
Zappia
V
V
D012437:Adenosylmethionine Decarboxylase / Q000302:isolation & purification* / Q000378:metabolism; D001105:Archaea / Q000201:enzymology*; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006358:Hot Temperature; D006882:Hydroxyapatites; D007202:Indicators and Reagents; D007700:Kinetics; D011773:Pyruvates / Q000032:analysis; D013816:Thermodynamics
D006882:Hydroxyapatites; D007202:Indicators and Reagents; D011773:Pyruvates; D017886:Durapatite; D012437:Adenosylmethionine Decarboxylase