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

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.

SUBMITTER: de Ruyck J

PROVIDER: S-EPMC3944699 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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

de Ruyck Jerome J Schubert Heidi L HL Janczak Matthew W MW Poulter C Dale CD

Acta crystallographica. Section F, Structural biology communications 20140219 Pt 3

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 ...[more]

PMID: 24598924

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