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BID peptides. More detailed biophysical and structural studies are in
progress.
A significant feature of the stapling approach is the supposed
enhanced cellular uptake conferred upon a peptide as a result of
stapling, however recent evidence has indicated that this is a
complex issue with experimental conditions playing a significant
role.36 Similarly in certain instances, whilst stapling does not
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its bioactive conformation in comparison to other modifications, it
can result in improved in cellulo PPI inhibition.11 The current
preliminary results thus contribute to the growing body of research
on peptide stapling; from a biophysical perspective, a modification
to any given peptide sequence e.g. the use of b-amino acids37,38
which tips the balance in favour of helix nucleation39,40 is what
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bioactive conformation. In contrast, cell uptake and function need
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In summary, we have shown that a-alkenyl substituted amino
acids act as effective amino acids for peptide stapling and demon-
strate that they can lead to comparable enhancement of proteolytic
stability, enhancement of helical propensity and similar efficiency
compared to peptides stapled with an a,a0-disubstituted variant
when applied to the BID BH3 helix and its inhibition of BCL-xL/
BAK as a model PPI. This modified amino acid is less sterically
encumbered than disubstituted amino acids and may facilitate
synthesis and therefore studies of stapled peptides. Our future
work will focus on structural and biophysical measurements on
this and other sequences in addition to more systematic studies on
cellular behaviour of constrained peptides.
This work was supported by the European Research Council
[ERC-StG-240324]. The authors wish to thank: Prof. W. D. Fairlie
(WEHI-Victoria) for the BCL-xL construct, Dr Alice Bartlett,
Dr Thomas Edwards and Hannah Kyle for performing protein
expression and purification, Dr Panchami Prabhakaran (BODIPY-
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Huscroft (HPLC), Tanya Marinko-Covell (HRMS) and Prof. Sheena
Radford for useful discussions.
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c
This journal is The Royal Society of Chemistry 2013
Chem. Commun., 2013, 49, 9131--9133 9133