Elongation factor T from Bacillus stearothermophilus and Escherichia coli. Purification and some properties of EF-Tu and EF-Ts from Bacillus stearothermophilus.
Homogeneous preparations of elongation factors EF-Tu and EF-Ts from Bacillus stearothermophilus have been obtained with specific activities of 20000 +/- 2000 and 500000 +/- 50000 units/mg, respectively. By dodecylsulphate-polyacrylamide gel electrophoresis the molecular weight of EF-Tu was found to be 49000 +/- 2000 and of EF-Ts 35500 +/- 1000. Nucleotide-free EF-Tu was prepared by using ITP as a GDP-binding-site-directed analogue. EF-Tu was shown to contain two sulphydryl groups, one reacting fast and one slowly with N-ethylmaleimide and 5,5'-dithio-bis(2-nitrobenzoic acid) under non-denaturing conditions. The same reagents were shown to react with the three sulphydryl groups of EF-Ts in the native state. The heat stabilities of EF-Tu and EF-Ts are reversed with respect to the Escherichia coli factors, EF-Tu being the more stable protein; even nucleotide-free EF-Tu is relatively stable with a half-life at room temperature of about 35 h.
D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D003460:Crystallization; D004355:Drug Stability; D004926:Escherichia coli / Q000032:analysis* / Q000378:metabolism; D005033:Ethylmaleimide; D001411:Geobacillus stearothermophilus / Q000032:analysis* / Q000378:metabolism; D006150:Guanine Nucleotides / Q000378:metabolism; D006358:Hot Temperature; D007292:Inosine Nucleotides / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D010445:Peptide Elongation Factors / Q000302:isolation & purification; D011485:Protein Binding; D013045:Species Specificity
D000596:Amino Acids; D006150:Guanine Nucleotides; D007292:Inosine Nucleotides; D010445:Peptide Elongation Factors; D005033:Ethylmaleimide