Overexpression, crystallization and preliminary X-ray crystallographic analysis of dihydrofolate reductase from bacteriophage T4

Acta Crystallogr D Biol Crystallogr. 2000 Jun;56(Pt 6):775-7. doi: 10.1107/s0907444900005266.

Abstract

Dihydrofolate reductase (DHFR) from bacteriophage T4 is a homodimer consisting of 193-residue subunits. It has been crystallized in the presence of the cofactor (NADPH) and an inhibitor (aminopterin) at 296 K using sodium chloride as precipitant. The crystals are tetragonal, belonging to the space group P4(1)22 (or P4(3)22), with unit-cell parameters a = b = 61.14, c = 123.23 A under cryogenic conditions. The asymmetric unit contains a single subunit, with a corresponding V(m) of 2.65 A(3) Da(-1) and a solvent content of 53. 6%. Native data have been collected from a crystal to 1.9 A resolution using synchrotron X-rays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage T4 / enzymology*
  • Bacteriophage T4 / genetics*
  • Crystallization
  • Crystallography, X-Ray
  • Data Collection
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Escherichia coli / virology
  • Gene Expression Regulation, Bacterial
  • Gene Expression Regulation, Viral
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Synchrotrons
  • Tetrahydrofolate Dehydrogenase / biosynthesis
  • Tetrahydrofolate Dehydrogenase / genetics*
  • Tetrahydrofolate Dehydrogenase / isolation & purification

Substances

  • Recombinant Proteins
  • Tetrahydrofolate Dehydrogenase