A lot of the interest in apoptosis mediation has focused on serine proteases, but research is at a very early stage so little is known about how these enzymes affect apoptosis
One of the most studied areas in life science research today is
More specifically, as a mechanism playing an essential
role in survival of an organism, apoptosis is providing an
opportunity for novel therapeutics, notably for anti-cancer
The key issue is identifying targets and then
understanding what effect they have in controlling apoptosis.
is in this aspect that Promega's ProteoLink in vitro
expression cloning system has been assisting one group at Cancer
A lot of the interest in apoptosis mediation has
focused on serine proteases, but research is at a very early
stage so little is known about how these enzymes affect
Miguel Martins's group at Cancer Research UK is
looking at one particular protease called Omi/HtrA2.
results have shown that this enzyme can process itself, but no
other substrates have been identified.
If other substrates can be
identified, Omi/HtrA2 will be implicated in a key stage in
In order to identify a substrate (or indeed several),
the entire cell proteome has to be screened.
there needs to be a facility to link proteomic information bank
to gene sequence data; but traditional methods (such as 2D gel
analysis or yeast two-hydrid screening) fail in this critical
In vitro translation, however, addresses this problem.
Promega's ProteoLink system provides a fast convenient
method that avoids the time-consuming steps of protein
purification, peptide sequencing and cDNA isolation.
adaptability of the method to a robotic platform enabled Miguel
Martins's group to rapidly and consistently process several
rounds of 96-well mini preps.
From the initial screen, eight
positive pools were detected which were further refined to a
single pool upon re-analysis with a lower protein loading (to
avoid non-specific digestion).
This pool was then analysed to
yield a single positive protein among the cDNA clones.
currently on-going to characterise this positive protein to see
if it fulfils the expected criteria for a Omi/HtrA2 target which,
in turn, will provide insight into the role this protease plays