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Purification, crystallization, and preliminary X‐ray studies of a bifunctional 5,10‐methenyl/methylene tetrahydrofolate cyclohydrolase/dehydrogenase from Escherichia coli
Author(s) -
Cheung Edwin,
D'Ari Linda,
Rabinowitz Jesse C.,
Dyer David H.,
Huang JieYu,
Stoddard Barry L.
Publication year - 1997
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/(sici)1097-0134(199702)27:2<322::aid-prot19>3.0.co;2-o
Subject(s) - bifunctional , methylene , chemistry , crystallography , monomer , crystallization , biochemistry , organic chemistry , catalysis , polymer
A bifunctional enzyme that catalyzes the conversion of formyltetrahydrofolate to methylene‐tetrahydrofolate (5,10‐methenyltetrahydrofolate cyclohydrolase and 5,10‐methylene tetrahydrofolate dehydrogenease), 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 = 64.5 Å b = 84.9 Å c = 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. © 1997 Wiley‐Liss, Inc.