Open AccessPurification, crystallization and preliminary crystallographic analysis of archaeal 6‐pyruvoyl tetrahydrobiopterin synthase homologue PH0634 from Pyrococcus horikoshii OT3
Author(s) -
Bagautdinov Bagautdin,
Sugahara Mitsuaki,
Kunishima Naoki
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106051578
Subject(s) - pyrococcus horikoshii , tetrahydrobiopterin , orthorhombic crystal system , crystallography , crystallization , trimer , crystal structure , space group , stereochemistry , chemistry , x ray crystallography , cofactor , biochemistry , enzyme , diffraction , physics , organic chemistry , dimer , optics
6‐Pyruvoyl tetrahydrobiopterin synthase (PTPS) catalyses the conversion of dihydroneopterin triphosphate to 6‐pyruvoyl tetrahydropterin, the second of the three enzymatic steps in the synthesis of tetrahydrobiopterin from GTP. PH0634, a 13.51 kDa archaeal PTPS homologue from Pyrococcus horikoshii OT3, was overexpressed as native and selenomethionine‐substituted protein and the purified protein was crystallized by the oil‐microbatch method at 295 K. X‐ray diffraction data were collected to 2.1 Å resolution from the native crystal using synchrotron radiation at 100 K. The crystal belongs to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 35.83, b = 95.71, c = 105.65 Å. Threefold noncrystallographic symmetry was identified from self‐rotation calculations. Assuming the presence of a trimer in the asymmetric unit, the solvent content is 45% ( V M = 2.24 Å 3 Da −1 ). The selenomethionine‐substituted crystal is isomorphous to the native crystal and diffracts X‐rays to 2.9 Å.