Expression, purification, crystallization and preliminary X-ray analysis of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase

Acta Crystallogr D Biol Crystallogr. 2001 Jan;57(Pt 1):150-2. doi: 10.1107/s0907444900014669.

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 A, alpha = beta = gamma = 90 degrees. The crystals belong to space group P2(1)2(1)2 and diffract to a minimum d spacing of 2.3 A 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, V(M) = 2.37 A(3) Da(-1)), with a solvent content of 48%.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Crystallization
  • Crystallography, X-Ray
  • Deoxyadenosines / chemistry*
  • Escherichia coli / enzymology
  • Molecular Sequence Data
  • N-Glycosyl Hydrolases / chemistry*
  • Thionucleosides / chemistry*

Substances

  • Deoxyadenosines
  • Thionucleosides
  • 5'-methylthioadenosine
  • N-Glycosyl Hydrolases
  • adenosylhomocysteine nucleosidase

Associated data

  • GENBANK/U24438