Primary structure and crystallization of orotate phosphoribosyltransferase from Salmonella typhimurium.
Orotate phosphoribosyltransferase (OPRTase; EC 2.4.2.10) catalyzes phosphoribosyl group transfer between alpha-D-5-phosphoribosyl-1-pyrophosphate and orotate to form orotidine-5'-monophosphate and pyrophosphate, the nucleotide-forming step in pyrimidine biosynthesis. It is one of ten PRTases that perform vital roles in de novo and salvage pathways for purine, pyrimidine and pyridine nucleotides. Although the PRTases are important drug targets, they are poorly understood mechanistically, and no three-dimensional structures exist. Here, we report the complete sequence of the Salmonella typhimurium pyrE gene and the deduced sequence of the OPRTase gene product. OPRTase forms tetragonal crystals from polyethylene glycol solutions; these crystals diffract to better than 2 A resolution, and are stable to radiation damage. The space group is P4(1)2(1)2 (or P4(3)2(1)2) with unit cell dimensions of a = b = 48.5 A, c = 210.5 A, and alpha = beta = gamma = 90 degrees. A crystalline form of the selenomethionine derivative of the protein is also reported.
Scapin
G
G
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
Sacchettini
J C
JC
Dessen
A
A
Bhatia
M
M
Grubmeyer
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009962:Orotate Phosphoribosyltransferase / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000235:genetics; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D009962:Orotate Phosphoribosyltransferase