PMID:11456491
| Citation |
Krab, IM, te Biesebeke, R, Bernardi, A and Parmeggiani, A (2001) Elongation factor Ts can act as a steric chaperone by increasing the solubility of nucleotide binding-impaired elongation factor-Tu. Biochemistry 40:8531-5 |
|---|---|
| Abstract |
Several elongation factor (EF) Tu mutants (T25A, H22Y/T25S, D80N, D138N) that have impaired nucleotide binding show decreased solubility on overexpression in the E. coli cell, an indication that they do not fold correctly. Moreover, EF-Tu[T25A] and EF-Tu[D80N] were shown to inhibit cell growth on expression, an effect attributed to their sequestration of EF-Ts [Krab, I. M., and Parmeggiani, A. (1999) J. Biol. Chem. 274, 11132--11138; Krab, I. M., and Parmeggiani, A. (1999) Biochemistry 38, 13035--13041]. We present here results showing that the co-overexpression of EF-Ts at a 1:1 ratio dramatically improves the solubility of mutant EF-Tu, although in the case of EF-Tu[D138N]--which cannot at all bind the nucleotides available in the cell--this is a slow process. Moreover, with co-overexpression of EF-Ts, the mentioned growth inhibition is relieved. We conclude that for the formation of a correct EF-Tu structure the nucleotide plays an important role as a "folding nucleus", and also that in its absence EF-Ts can act as a folding template or steric chaperone for the correct folding of EF-Tu. |
| Links | |
| Keywords |
Escherichia coli/genetics; Glutathione Transferase/genetics; Growth Inhibitors/chemistry; Guanine Nucleotides/chemistry; Molecular Chaperones/biosynthesis; Molecular Chaperones/chemistry; Molecular Chaperones/genetics; Mutagenesis, Site-Directed; Peptide Elongation Factor Tu/biosynthesis; Peptide Elongation Factor Tu/chemistry; Peptide Elongation Factor Tu/genetics; Peptide Elongation Factors/biosynthesis; Peptide Elongation Factors/chemistry; Peptide Elongation Factors/genetics; Plasmids/biosynthesis; Protein Binding; Protein Folding; Recombinant Fusion Proteins/biosynthesis; Ribonucleosides/genetics; Solubility |
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