Streptomyces aspartate transcarbamoylase is a dodecamer with dihydroorotase activity.
Aspartate transcarbamoylase (ATCase) was purified from Streptomyces griseus. The enzyme is a dodecamer with a molecular mass of approximately 450 kDa. The holoenzyme is a complex of ATCase and active dihydroorotase (DHOase) subunits. The ATCase and DHOase activities co-purify after gel filtration and ion-exchange chromatography. Denaturing gel electrophoresis separates the holoenzyme into a 38-kDa ATCase polypeptide and a 47-kDa DHOase polypeptide. The holoenzyme retained ATCase and DHOase activity after being heated to 65 degrees C for 5 min, but after storage at 4 degrees C for 24 hours lost ATCase activity. Previously, the Pseudomonas putida Class A ATCase was defined by Schurr et al. (J Bacteriol 177, 1751-1759) as requiring an inactive DHOase to be functional. Here, we show that an active DHOase is part of the dodecameric ATCase/DHOase complex in Streptomyces. To distinguish those Class A ATCases with active DHOases from those with degenerate DHOases, we suggest the subdivision, Class A(1), for the former and Class A(2) for the latter.
Hughes
L E
LE
Department of Biological Sciences, University of North Texas, Denton, Texas 76203, USA.
Hooshdaran
M Z
MZ
O'Donovan
G A
GA
D001221:Aspartate Carbamoyltransferase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015153:Blotting, Western; D004080:Dihydroorotase / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D020035:Holoenzymes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D013305:Streptomyces griseus / Q000201:enzymology* / Q000254:growth & development
D020035:Holoenzymes; D001221:Aspartate Carbamoyltransferase; D004080:Dihydroorotase