Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis.
Phosphopantetheine adenylyltransferase, an essential enzyme in the coenzyme A biosynthetic pathway, catalyzes the reversible transfer of an adenylyl group from ATP to 4'-phosphopantetheine, yielding 3'-dephospho-CoA and pyrophosphate. Enterococcus faecalis PPAT has been overexpressed in Escherichia coli as a fusion with a C-terminal purification tag and crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium HEPES pH 7.5, 0.8 M sodium dihydrogen phosphate and 0.8 M potassium dihydrogen phosphate. X-ray diffraction data were collected to 2.70 A at 100 K. The crystals belong to the primitive tetragonal space group P4(1) (or P4(3)), with unit-cell parameters a = b = 160.81, c = 225.68 A. Four copies of the hexameric molecule are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (V(M)) of 3.08 A(3) Da(-1) and a solvent content of 60.1%.
Kang
Ji Yong
JY
Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul 151-742, South Korea.
Lee
Hyung Ho
HH
Yoon
Hye Jin
HJ
Kim
Hyoun Sook
HS
Suh
Se Won
SW
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D013293:Enterococcus faecalis / Q000201:enzymology*; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D012542:Scattering, Radiation; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D017931:DNA Primers; D009713:Nucleotidyltransferases; C022315:pantetheine-phosphate adenylyltransferase