Crystallization and preliminary X-ray crystallographic analysis of quinolinate phosphoribosyltransferase of Helicobacter pylori.
Quinolinic acid phosphoribosyltransferase (NadC; EC 2.4.2.19) is the key enzyme of NAD(+) biosynthesis in both prokaryotes and eukaryotes. NadC catalyzes the decarboxylation of quinolinic acid (QA) to produce nicotinic acid mononucleotide (NAMN), an intermediate in NAD synthesis. NadCs of Helicobacter pylori appeared to be a hexamer during the purification procedure. Three different complexes of NadC, with QA, NAMN and phthalic acid (PA), an analogue of QA, were crystallized at 294 +/- 1 K using the hanging-drop vapour-diffusion method. The QA complex crystal was found to belong to space group P4(1)2(1)2, with unit-cell parameters a = b = 148.8, c = 145.7 A, alpha = beta = gamma = 90 degrees. Diffraction data were collected from the NadC-substrate and NadC-substrate analogue complexes to resolutions of 2.3 A (QA), 2.8 A (PA) and 3.3 A (NAMN) using synchrotron X-ray radiation.
Kim
Mun-Kyoung
MK
Department of Life Science, Kwangju Institute of Science and Technology, Gwangju 500-712, South Korea.
Kim
Yun Sik
YS
Rho
Seong-Hwan
SH
Im
Young Jun
YJ
Lee
Jun Hyuck
JH
Kang
Gil Bu
GB
Eom
Soo Hyun
SH
D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D016480:Helicobacter pylori / Q000201:enzymology*; D009537:Nicotinamide Mononucleotide / Q000031:analogs & derivatives* / Q000737:chemistry; D010430:Pentosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D010795:Phthalic Acids / Q000737:chemistry; D011485:Protein Binding; D017378:Quinolinic Acid / Q000737:chemistry; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating)
D010795:Phthalic Acids; D009537:Nicotinamide Mononucleotide; C002953:nicotinate mononucleotide; C032279:phthalic acid; D010430:Pentosyltransferases; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating); D017378:Quinolinic Acid