Work

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

Year: 2014

Type: article

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.

Source: Acta Crystallographica Section F Structural Biology Communications

DOI: 10.1107/s2053230x14002143

Authors: Jerome de Ruyck, Heidi L. Schubert, Matthew W. Janczak, C. Dale Poulter

Institutions: University of Namur, University of Utah

Language: English

Topic: Enzyme Structure and Function

Subfield: Materials Chemistry

Field: Materials Science

Domain: Physical Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Awards: G4212847572, Studies of Isoprenoid Biosynthesis, A Synchrotron Radiation Structural Biology Resource