Isolation, crystallisation, and preliminary X-ray analysis of the bovine mitochondrial EF-Tu:GDP and EF-Tu:EF-Ts complexes

Biochim Biophys Acta. 2002 Dec 16;1601(2):172-7. doi: 10.1016/s1570-9639(02)00460-0.

Abstract

Previous studies have shown that when bovine mitochondrial elongation factor Ts (EF-Ts) is expressed in Escherichia coli, it forms a tightly associated complex with E. coli elongation factor Tu (EF-Tu). In contrast to earlier experiments, purification of free mitochondrial EF-Ts was accomplished under nondenaturing conditions since only about 60% of the expressed EF-Ts copurified with E. coli EF-Tu. The bovine mitochondrial EF-Tu:GDP complex, the homologous mitochondrial EF-Tu:EF-Ts complex, and the heterologous E. coli/mitochondrial EF-Tu:EF-Ts complex were isolated and crystallised. The crystals of the EF-Tu:GDP complex diffract to 1.94 A and belong to space group P2(1) with cell parameters a=59.09 A, b=119.78 A, c=128.89 A and beta=96.978 degrees. The crystals of the homologous mitochondrial EF-Tu:EF-Ts complex diffract to 4 A and belong to space group C2 with cell parameters a=157.7 A, b=151.9 A, c=156.9 A, and beta=108.96 degrees.

Publication types

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

MeSH terms

  • Animals
  • Cattle
  • Cells, Cultured
  • Chromatography, Ion Exchange
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Guanosine Diphosphate / chemistry*
  • Guanosine Diphosphate / isolation & purification
  • Mitochondria / metabolism*
  • Peptide Elongation Factor Tu / chemistry*
  • Peptide Elongation Factor Tu / isolation & purification
  • Peptide Elongation Factors / chemistry*
  • Peptide Elongation Factors / isolation & purification

Substances

  • Peptide Elongation Factors
  • elongation factor Ts
  • Guanosine Diphosphate
  • Peptide Elongation Factor Tu