Expression, purification, crystallization and preliminary x-ray analysis of escherichia coli 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidase

J. E. Lee, K. A. Cornell, Michael Riscoe, P. Lynne Howell

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A recombinant form of Escherichia coli 5'-methylthioadenosine/ S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 Å, a = β = γ = 90°. The crystals belong to space group P21212 and diffract to a minimum d spacing of 2.3 Å on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, VM = 2.37 Å3 Da-1), with a solvent content of 48%.

Original languageEnglish (US)
Pages (from-to)150-152
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume57
Issue number1
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

adenosylhomocysteine nucleosidase
x ray analysis
Diamond
Escherichia
Crystallization
purification
X-Ray Diffraction
Escherichia coli
Purification
Electrodes
X-Rays
crystallization
X rays
Crystals
crystals
wedges
X ray diffraction analysis
homogeneity
Anodes
anodes

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

@article{de5b46df8cee4e3da886c12c15dd5092,
title = "Expression, purification, crystallization and preliminary x-ray analysis of escherichia coli 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidase",
abstract = "A recombinant form of Escherichia coli 5'-methylthioadenosine/ S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 {\AA}, a = β = γ = 90°. The crystals belong to space group P21212 and diffract to a minimum d spacing of 2.3 {\AA} on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, VM = 2.37 {\AA}3 Da-1), with a solvent content of 48{\%}.",
author = "Lee, {J. E.} and Cornell, {K. A.} and Michael Riscoe and {Lynne Howell}, P.",
year = "2001",
doi = "10.1107/S0907444900014669",
language = "English (US)",
volume = "57",
pages = "150--152",
journal = "Acta Crystallographica Section D: Structural Biology",
issn = "0907-4449",
publisher = "John Wiley and Sons Inc.",
number = "1",

}

TY - JOUR

T1 - Expression, purification, crystallization and preliminary x-ray analysis of escherichia coli 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidase

AU - Lee, J. E.

AU - Cornell, K. A.

AU - Riscoe, Michael

AU - Lynne Howell, P.

PY - 2001

Y1 - 2001

N2 - A recombinant form of Escherichia coli 5'-methylthioadenosine/ S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 Å, a = β = γ = 90°. The crystals belong to space group P21212 and diffract to a minimum d spacing of 2.3 Å on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, VM = 2.37 Å3 Da-1), with a solvent content of 48%.

AB - A recombinant form of Escherichia coli 5'-methylthioadenosine/ S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 Å, a = β = γ = 90°. The crystals belong to space group P21212 and diffract to a minimum d spacing of 2.3 Å on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, VM = 2.37 Å3 Da-1), with a solvent content of 48%.

UR - http://www.scopus.com/inward/record.url?scp=0035180468&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035180468&partnerID=8YFLogxK

U2 - 10.1107/S0907444900014669

DO - 10.1107/S0907444900014669

M3 - Article

C2 - 11134941

AN - SCOPUS:0035180468

VL - 57

SP - 150

EP - 152

JO - Acta Crystallographica Section D: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

SN - 0907-4449

IS - 1

ER -