Isolation, crystallisation, and preliminary X-ray analysis of the bovine mitochondrial EF-Tu:GDP and EF-Tu:EF-Ts complexes.
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.
Karring
Henrik
H
Department of Molecular and Structural Biology, Aarhus University, Gustav Wiedsvej 10c, Arhus C DK-8000, Denmark. kar@biobase.dk
Andersen
Gregers R
GR
Thirup
Søren S
SS
Nyborg
Jens
J
Spremulli
Linda L
LL
Clark
Brian F C
BF
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D006153:Guanosine Diphosphate / Q000737:chemistry* / Q000302:isolation & purification; D008928:Mitochondria / Q000378:metabolism*; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000302:isolation & purification; D010445:Peptide Elongation Factors / Q000737:chemistry* / Q000302:isolation & purification
D010445:Peptide Elongation Factors; C023512:elongation factor Ts; D006153:Guanosine Diphosphate; D010444:Peptide Elongation Factor Tu