Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8. 1. Purification and some properties of the purified factors.
Polypeptide chain elongation factors have been purified from an extreme thermophile, Thermus thermophilus HB8. By chromatography on a DEAE-Sephadex column, the factors were separated into two peaks; peak I contained a complex of EF-Tu and EF-Ts, while peak II was composed of EF-Tu.gdp and EF-G. These factors were subsequently purified to homogeneous states and crystallized. The EF-Tu . EF-Ts complex could be resolved into EF-Tu and EF-Ts by chromatography on a Sephadex G-200 column in the presence of 8 M guanidine-HCl. The complex could be reconstituted from EF-Tu and the renatured EF-Ts. No immunological cross-reaction was detected between EF-Tu, EF-Ts, and EF-G from T. thermophilus and the antibodies to their corresponding Escherichia coli factors. The molecular weight of EF-Tu . GDP determined by sedimentation equilibrium and sodium dodecylsulfate/polyacrylamide gel electrophoresis was 49000 and 51000 respectively. On the other hand, the molecular weight of EF-Ts was estimated as 27000 and 64000, respectively, by sodium dodecylsulfate/polyacrylamide gel electrophoresis and Sephadex gel filtration, suggesting that the protein existed probably as a dimer. The molecular weight of the EF-Tu . EF-Ts complex determined by sedimentation equilibrium and by gel filtration, was 142000 and 220000, respectively. Since the molar ratio of EF-Tu to EF-Ts in the EF-Tu . EF-Ts complex was one to one, it was suggested that the complex was composed of 2 mol each of EF-Tu and EF-Ts. The molecular weight of EF-G was estimated as 85000, 80000 and 78000 by equilibrium centrifugation, gel filtration, and sodium dodecylsulfate/polyacrylamide gel electrophoresis respectively.
D003460:Crystallization; D008722:Methods; D008970:Molecular Weight; D010445:Peptide Elongation Factors / Q000276:immunology / Q000302:isolation & purification*; D011487:Protein Conformation; D013824:Thermus / Q000378:metabolism*
D010445:Peptide Elongation Factors