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. 2014 Mar;70(Pt 3):347-9.
doi: 10.1107/S2053230X14002143. Epub 2014 Feb 19.

Tetartohedral twinning in IDI-2 from Thermus thermophilus: crystallization under anaerobic conditions

Jerome de Ruyck  1 , Heidi L Schubert  2 , Matthew W Janczak  3 , C Dale Poulter  3
Affiliations

Tetartohedral twinning in IDI-2 from Thermus thermophilus: crystallization under anaerobic conditions

Jerome de Ruyck et al. Acta Crystallogr F Struct Biol Commun. 2014 Mar.

Abstract

Type-2 isopentenyl diphosphate isomerase (IDI-2) is a key flavoprotein involved in the biosynthesis of isoprenoids. Since fully reduced flavin mononucleotide (FMNH2) 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 P6322, with unit-cell parameters a = b = 133.3, c = 172.9 Å.

Keywords: IDI-2; anaerobic; flavoprotein; isopentenyl diphosphate isomerase; isoprenoid; twinning.

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Figures

Figure 1
Figure 1
Crystals of colourless tt-IDI-2.
Figure 2
Figure 2
Diffraction patterns of (a) native and (b) soaked crystals. These patterns were taken on an in-house diffractometer prior to synchrotron data collection.
See this image and copyright information in PMC

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