Purification and characterization of elongation factor G from bovine liver mitochondria.
The mitochondrial protein synthesis translocase elongation factor Gmt (EF-Gmt) from bovine liver has been purified to greater than 90% homogeneity by a combination of conventional gravity and high performance liquid chromatography. The purification scheme results in an approximate overall 14,000-fold purification with 2% total recovery of EF-Gmt activity. Gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicate that the mitochondrial factor is a single polypeptide with a molecular weight of 80,000. EF-Gmt displays similar levels of activity on its homologous mitochondrial ribosomes and on Escherichia coli ribosomes. The mitochondrial translocase is sensitive to temperatures above 37 degrees C, but the factor is partially protected from heat inactivation in the presence of GTP or GDP. The activity of EF-Gmt is inhibited by treatment of the factor with N-ethylmaleimide. In contrast to all other translocases tested to date, EF-Gmt is completely resistant to the inhibiting effect of fusidic acid when tested on its homologous ribosomes. It displays weak sensitivity to this antibiotic when assayed in the presence of heterologous E. coli ribosomes.
Chung
H K
HK
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290.
Spremulli
L L
LL
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005672:Fusidic Acid / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008930:Mitochondria, Liver / Q000378:metabolism*; D008970:Molecular Weight; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors / Q000302:isolation & purification* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D006153:Guanosine Diphosphate; D005672:Fusidic Acid; D006160:Guanosine Triphosphate