Conformational Analysis of the Host-Defense Peptides
Pseudhymenochirin-1Pb and -2Pa and Design of Analogues with Insulin-Releasing
Activities and Reduced Toxicities
posted on 2015-12-24, 00:00authored byGiorgia Manzo, Mariano
Andrea Scorciapino, Dinesh Srinivasan, Samir Attoub, Maria Luisa Mangoni, Andrea C. Rinaldi, Mariano Casu, Peter
R. Flatt, J. Michael Conlon
Pseudhymenochirin-1Pb (Ps-1Pb; IKIPSFFRNILKKVGKEAVSLIAGALKQS)
and pseudhymenochirin-2Pa (Ps-2Pa; GIFPIFAKLLGKVIKVASSLISKGRTE)
are amphibian peptides with broad spectrum antimicrobial activities
and cytotoxicity against mammalian cells. In the membrane-mimetic
solvent 50% (v/v) trifluoroethanol–H2O, both peptides
adopt a well-defined α-helical conformation that extends over
almost all the sequence and incorporates a flexible bend. Both peptides
significantly (p < 0.05) stimulate the rate of
release of insulin from BRIN-BD11 clonal β-cells at concentrations
≥ 0.1 nM but produce loss of integrity of the plasma membrane
at concentrations ≥ 1 μM. Increasing cationicity by the
substitution Glu17 → l-Lys in Ps-1Pb and
Glu27 → l-Lys in Ps-2Pa generates analogues
with increased cytotoxicity and reduced insulin-releasing potency.
In contrast, the analogues [R8r]Ps-1Pb and [K8k,K19k]Ps-2Pa, incorporating d-amino acid residues to destabilize the α-helical domains,
retain potent insulin-releasing activity but are nontoxic to BRIN-BD11
cells at concentrations of 3 μM. [R8r]Ps-1Pb produces a significant
increase in insulin release rate at 0.3 nM and [K8k,K19k]Ps-2Pa at
0.01 nM. Both analogues show low hemolytic activity (IC50 > 100 μM) but retain broad-spectrum antimicrobial activity
and remain cytotoxic to a range of human tumor cell lines, albeit
with lower potency than the naturally occurring peptides. These analogues
show potential for development into agents for type 2 diabetes therapy.