Mutation of the conserved Gly83 and Gly94 in Escherichia coli elongation factor Tu. Indication of structural pivots.

I V Kjaersgård, Charlotte Rohde Knudsen, O Wiborg

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    Abstract

    Elongation factor Tu from Escherichia coli cycles between an active conformation where GTP is bound, and an inactive conformation where GDP is bound. Between the two conformations, elongation factor Tu undergoes major structural changes. The aim of this work has been to reveal the role of two very well conserved glycine residues, Gly83 and Gly94, in the switch mechanism. Gly83 has been mutated alone or in combination with Gly94, both glycine residues being mutated to alanine. Enzymic characterisation of the two mutants have shown that they have an altered nucleotide affinity, a decrease in aminoacyl-tRNA affinity, an increase in intrinsic GTP hydrolysis, different behaviours in effector stimulation of the intrinsic GTPase activity, and that they are completely unable to sustain poly(Phe) synthesis in an in-vitro poly(U)-directed system. Our results indicates that particularly Gly83 is an important pivot point in elongation factor-Tu.
    Udgivelsesdato: 1995-Feb-15
    Original languageEnglish
    JournalF E B S Journal
    Volume228
    Issue1
    Pages (from-to)184-90
    Number of pages6
    ISSN1742-464X
    Publication statusPublished - 1995

    Keywords

    • Base Sequence
    • GTP Phosphohydrolase-Linked Elongation Factors
    • Guanosine Diphosphate
    • Guanosine Triphosphate
    • Molecular Sequence Data
    • Mutation
    • Peptide Elongation Factor Tu
    • Protein Biosynthesis
    • Structure-Activity Relationship

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