Purification, crystallization, and preliminary X-ray studies of a bifunctional 5,10-methenyl/methylene tetrahydrofolate cyclohydrolase/dehydrogenase from Escherichia coli
✍ Scribed by Edwin Cheung; Linda D'Ari; Jesse C. Rabinowitz; David H. Dyer; Jie-Yu Huang; Barry L. Stoddard
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 32 KB
- Volume
- 27
- Category
- Article
- ISSN
- 0887-3585
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✦ Synopsis
A bifunctional enzyme that catalyzes the conversion of formyltetrahydrofolate to methylene-tetrahydrofolate (5,10methenyltetrahydrofolate cyclohydrolase and 5,10-methylene tetrahydrofolate dehydrogenase), has been subcloned from a cDNA library, purified to homogeneity, and crystallized. The crystals belong to space group I222, with unit cell dimensions of a 5 64.5 Å, b 5 84.9 Å, c 5 146.1 Å. The crystal unit cell and diffraction is consistent with an asymmetric unit consisting of the enzyme monomer, and a specific volume of the unit cell of 3.2 Å 3 /Da. The crystals diffract to at least 2.8 Å resolution after flash-cooling, when using a rotating anode x-ray source and an RAXIS image plate detector. A 2.56 Å resolution native data set has been collected at beamline X12-C at the NSLS.