Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP +

Qun Wan, Andrey Y. Kovalevsky, Mark A Wilson, Brad C. Bennett, Paul Langan, Chris Dealwis

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A crystal of Escherichia coli dihydrofolate reductase (ecDHFR) complexed with folate and NADP + of 4 × 1.3 × 0.7 mm (3.6 mm 3 ) 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.

Original languageEnglish (US)
Pages (from-to)814-818
Number of pages5
JournalActa Crystallographica Section F:Structural Biology Communications
Volume70
Issue number6
DOIs
StatePublished - Jan 1 2014

Fingerprint

Tetrahydrofolate Dehydrogenase
Neutrons
Escherichia
NADP
Folic Acid
Escherichia coli
high flux isotope reactors
Joints
X-Rays
Neutron Diffraction
proteins
neutrons
X rays
Crystals
Protonation
Diffractometers
Neutron diffraction
diffractometers
Isotopes
crystals

Keywords

  • Escherichia coli
  • dihydrofolate reductase
  • protonation state

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP + . / Wan, Qun; Kovalevsky, Andrey Y.; Wilson, Mark A; Bennett, Brad C.; Langan, Paul; Dealwis, Chris.

In: Acta Crystallographica Section F:Structural Biology Communications, Vol. 70, No. 6, 01.01.2014, p. 814-818.

Research output: Contribution to journalArticle

@article{824b18233b664eb88075b9f100a66f01,
title = "Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP +",
abstract = "A crystal of Escherichia coli dihydrofolate reductase (ecDHFR) complexed with folate and NADP + of 4 × 1.3 × 0.7 mm (3.6 mm 3 ) in size was obtained by sequential application of microseeding and macroseeding. A neutron diffraction data set was collected to 2.0 {\AA} resolution using the IMAGINE diffractometer at the High Flux Isotope Reactor within Oak Ridge National Laboratory. A 1.6 {\AA} 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 = "Escherichia coli, dihydrofolate reductase, protonation state",
author = "Qun Wan and Kovalevsky, {Andrey Y.} and Wilson, {Mark A} and Bennett, {Brad C.} and Paul Langan and Chris Dealwis",
year = "2014",
month = "1",
day = "1",
doi = "10.1107/S2053230X1400942X",
language = "English (US)",
volume = "70",
pages = "814--818",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",
number = "6",

}

TY - JOUR

T1 - Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP +

AU - Wan, Qun

AU - Kovalevsky, Andrey Y.

AU - Wilson, Mark A

AU - Bennett, Brad C.

AU - Langan, Paul

AU - Dealwis, Chris

PY - 2014/1/1

Y1 - 2014/1/1

N2 - A crystal of Escherichia coli dihydrofolate reductase (ecDHFR) complexed with folate and NADP + of 4 × 1.3 × 0.7 mm (3.6 mm 3 ) 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.

AB - A crystal of Escherichia coli dihydrofolate reductase (ecDHFR) complexed with folate and NADP + of 4 × 1.3 × 0.7 mm (3.6 mm 3 ) 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.

KW - Escherichia coli

KW - dihydrofolate reductase

KW - protonation state

UR - http://www.scopus.com/inward/record.url?scp=84905496762&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84905496762&partnerID=8YFLogxK

U2 - 10.1107/S2053230X1400942X

DO - 10.1107/S2053230X1400942X

M3 - Article

VL - 70

SP - 814

EP - 818

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

IS - 6

ER -