Open AccessOverexpression, crystallization and preliminary X‐ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis
Author(s) -
Kang Ji Yong,
Lee Hyung Ho,
Yoon Hye Jin,
Kim Hyoun Sook,
Suh Se Won
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106041108
Subject(s) - pyrophosphate , crystallization , enterococcus faecalis , chemistry , crystallography , monoclinic crystal system , potassium , sodium , escherichia coli , tetragonal crystal system , transferase , stereochemistry , crystal structure , enzyme , biochemistry , organic chemistry , gene
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 Å at 100 K. The crystals belong to the primitive tetragonal space group P 4 1 (or P 4 3 ), with unit‐cell parameters a = b = 160.81, c = 225.68 Å. 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 Å 3 Da −1 and a solvent content of 60.1%.