Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP+
Open Access
- 25 May 2014
- journal article
- crystallization communications
- Published by International Union of Crystallography (IUCr) in Acta Crystallographica Section F Structural Biology Communications
- Vol. 70 (6) , 814-818
Abstract
A crystal ofEscherichia colidihydrofolate reductase (ecDHFR) complexed with folate and NADP+of 4 × 1.3 × 0.7 mm (3.6 mm3) in size was obtained by sequential application of microseeding and macroseeding. A neutron diffraction data set was collected to 2.0 Å resolution using the IMAGINE diffractometer at the High Flux Isotope Reactor within Oak Ridge National Laboratory. A 1.6 Å resolution X-ray data set was also collected from a smaller crystal at room temperature. The neutron and X-ray data were used together for joint refinement of the ecDHFR–folate–NADP+ternary-complex structure in order to examine the protonation state, protein dynamics and solvent structure of the complex, furthering understanding of the catalytic mechanism.Keywords
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