Premium
Tetartohedral twinning in IDI‐2 from Thermus thermophilus : crystallization under anaerobic conditions
Author(s) -
de Ruyck Jerome,
Schubert Heidi L.,
Janczak Matthew W.,
Poulter C. Dale
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14002143
Subject(s) - thermus thermophilus , flavin mononucleotide , isomerase , cofactor , flavoprotein , anaerobic exercise , crystallization , chemistry , stereochemistry , crystallography , enzyme , biochemistry , biology , organic chemistry , escherichia coli , physiology , gene
Type‐2 isopentenyl diphosphate isomerase (IDI‐2) is a key flavoprotein involved in the biosynthesis of isoprenoids. Since fully reduced flavin mononucleotide (FMNH 2 ) is needed for activity, it was decided to crystallize the enzyme under anaerobic conditions in order to understand how this reduced cofactor binds within the active site and interacts with the substrate isopentenyl diphosphate (IPP). In this study, the protein was expressed and purified under aerobic conditions and then reduced and crystallized under anaerobic conditions. Crystals grown by the sitting‐drop vapour‐diffusion method and then soaked with IPP diffracted to 2.1 Å resolution and belonged to the hexagonal space group P 6 3 22, with unit‐cell parameters a = b = 133.3, c = 172.9 Å.