Adsorption properties of new antifungal peptides derived

from Silybum marianum defensins.

Iturralde, Micaela; Bracho Oliveros, JP.; Valdivia Pérez, J.A.; Guzmán, F.; Maté, S.; Vairo Cavalli, S.E.;
Fanani, M.L.

Natural antimicrobial peptides (AMPs) provide a valuable basis for the development of
new AMPs with enhanced features through rational design. We have designed and
synthesized peptides from the ɣ-core and α-core motifs of the defensins DefSm2-D and
DefSm3-D (Silybum marianum), which have different amino acid residues in the α-core
region. The peptide SmAPɣ27-44 sequence is common to both defensins, while SmAP α1-21
and SmAP3α1-21 sequences are specific to each defensin. SmAP ɣ27-44 and SmAPα1-21 have
antifungal activity against Fusarium graminearum.
Large unilamellar liposomes (LUVs) composed of PC/PE/PA/Erg 5:4:1:2 (w/w/w/w)
were used as models of fungal plasma membrane. PA is a lipid present in filamentous
fungi and has a relevant role in the mechanism of action of various AMPs. For example,
for the binding of MtDef4, a peptide with well-known antifungal activity against F.
graminearum.
LUVs were analysed by Z potential and size measurements by dynamic light scattering
against different peptide concentrations. The binding of AMPs to LUVs was assessed
using red edge excitation shift (REES) experiments to evaluate the environment around
the tryptophan (Trp) residue.
Our results indicate that the behaviour of the peptides against LUVs presented
considerable variations between them. SmAP3 α1-21 has a minimum inhibitory
concentration (MIC) against F. graminearum greater than 100 M and shows poor
adsorption to liposomes. On the other hand, the two most active peptides SmAP α1-21
(MIC= 32 M) and SmAPɣ27-44 (MIC= 20 M) show different mechanisms of action. The
first one strongly induces the fusion of liposomes, showing an insertion of its Trp moiety
into the hydrophobic interior of the membrane, while the second is characterized by an
adsorption that results in the neutralization of the surface charge.

Acknowledgements. PICT 2019-2598, PICT 2019-0332 and Federal Mobility Program

FONCyT 2022.