Project Details


We have recently identified a new class of protein kinases, which
are structurally and evolutionarily unrelated to members of the
eukaryotic protein kinase superfamily. The prototype member of
this class is Ca2+/calmodulin-dependent elongation factor-2
kinase (eEF-2 kinase), a ubiquitous protein kinase present in
various eukaryotic organisms. The major function of eEF-2 kinase
is to phosphorylate and inactivate eEF-2, and thus, regulate the
elongation phase of protein synthesis. Preliminary evidence
suggests that eEF-2 kinase has a novel type of catalytic domain,
and utilizes a novel mechanism of substrate recognition.

The aim of this proposal is to identify and characterize the
functional domains of eEF-2 kinase, to study its mechanism of
substrate recognition, and to reveal its specific physiological

In vitro mutagenesis of human eEF-2 kinase will be performed to
identify and characterize the catalytic and calmodulin-binding
domains. We will also study the mechanism of substrate
recognition by eEF-2 kinase. An oriented peptide library will
be screened to identify the consensus sequence recognized by eEF-
2 kinase. Next, we will test the hypothesis that an alpha-
helical conformation of the peptide at the phosphorylation site
of the substrate is required for recognition by eEF-2 kinase.
Peptides mimicking phosphorylation sites with varying degrees of
alpha-helicity will be synthesized and tested for their ability
to undergo phosphorylation.

To identify other potential substrates for eEF-2 kinase, two
approaches will be used: a solid-phase phosphorylation expression
screening method and the yeast two-hybrid system.

To reveal the exact physiological function of eEF-2 kinase,
Caenorhabditis elegans will be used. We will analyze the effect
of eEF-2 kinase gene inactivation on phenotype and overall
developmental pattern using three different approaches: Tc1
mutagenesis, deletion mutagenesis with rescue by plasmid
transgenics, and an antisense approach.

This work will reveal the structural organization and function of
a new class of protein kinases, which will provide new and
important information about the mechanism of protein
Effective start/end date1/1/9912/31/05


  • National Institute of General Medical Sciences: $254,570.00
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences: $247,250.00
  • National Institute of General Medical Sciences: $262,113.00
  • National Institute of General Medical Sciences: $252,389.00


  • Catalysis


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