Purification, crystallization and preliminary X-ray diffraction study on pyrimidine nucleoside phosphorylase TTHA1771 from Thermus thermophilus HB8.
Pyrimidine nucleoside phosphorylase (PYNP) catalyzes the reversible phosphorolysis of pyrimidines in the nucleotide-synthesis salvage pathway. In order to study the structure-thermostability relationship of this enzyme, PYNP from the extreme thermophile Thermus thermophilus HB8 (TTHA1771) has been cloned, overexpressed and purified. The TTHA1771 protein was crystallized at 291 K using the oil-microbatch method with PEG 4000 as a precipitant. A native data set was collected to 1.8 A resolution using synchrotron radiation. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 58.83, b = 76.23, c = 103.86 A, beta = 91.3 degrees.
Shimizu
Katsumi
K
Advanced Protein Crystallography Research Group, RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cyo, Sayo-gun, Hyogo 679-5148, Japan.
Kunishima
Naoki
N
D003460:Crystallization; D018360:Crystallography, X-Ray; D010430:Pentosyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D057845:Pyrimidine Phosphorylases; D016963:Thermus thermophilus / Q000201:enzymology*
D010430:Pentosyltransferases; D057845:Pyrimidine Phosphorylases