Participation of the overproduced elongation factor Tu fromThermus thermophilusin protein biosynthesis ofEscherichia coli

Conformational Change of Elongation Factor Tu (EF-Tu) Induced by Antibiotic Binding

“…The EF-Tu from this organism shares 82% sequence identity with the protein from Escherichia coli and has been shown to be similarly sensitive to kirromycin (16). The crystal structure presented here nicely explains the mutations reported to confer resistance to EF-Tu against the antibiotic (17)(18)(19)(20).…”

Site‐Directed Mutagenesis of Thermus thermophilus Elongation Factor Tu

“…This system is suited for studying the activity of EFTu variants in vivo. Since 7: thermophilus EF-Tu binds kirromycin and interacts with E. coli ribosomes, it also causes kirromycin sensitivity when produced in a kirromycin-resistant E. coli strain (Zeidler et al, 1993). EFTu show a similar effect (Fig.…”

“…Growth of E. coli strains on kirromycin was carried out as reported (Zeidler et al, 1993). The E. coli strain LBE2012/ pREP4 was transformed with the expression vectors pEFTulO, pEFTuH85G, pEFTuH85Q or pEFTuH85L.…”

Bacterial elongation factors EF-Tu, their mutants, chimeric forms, and domains: Isolation and purification

“…were reported: passage through a French press [23,60,77], grinding of cells with twice bacteria mass of alumina (Al 2 O 3 ) [42,44], lysis with lysozyme (EDTA, sodium deoxycholate, DNase I treatment) [32,54,59,62,71,78], etc.…”

“…For example, recombinant Thermus thermophilus, Thermus aquaticus, and Thermotoga maritima EF-Tus (all of thermophilic organisms) overproduced in E. coli cells, remain all active after heating at 65 • C when EcEF-Tu is completely denatured and precipitated and can be quantitatively removed with most other E. coli proteins simply by centrifugation [23,[40][41][42][43][44]. Beside the purification of EF-Tu, Blank et al [43] also described purification of EF-Ts and EF-G from T. thermophilus by this heat treatment.…”