Short oligoalanine helical peptides for supramolecular nanopore assembly and protein cytosolic delivery
In this work we report a rational design strategy for the identification of new peptide prototypes for the non-disruptive supramolecular permeation of membranes and the transport of different macromolecular giant cargos. The approach targets a maximal enhancement of helicity in the presence of membranes with sequences bearing the minimal number of cationic and hydrophobic moieties. The here reported folding enhancement in membranes allowed the selective non-lytic translocation of different macromolecular cargos including giant proteins. The transport of different high molecular weight polymers and functional proteins was demonstrated in vesicles and in cells with excellent efficiency and optimal viability. As a proof of concept, functional monoclonal antibodies were transported for the first time into different cell lines and cornea tissues by exploiting the helical control of a short peptide sequence. This work introduces a rational design strategy that can be employed to minimize the number of charges and hydrophobic residues of short peptide carriers to achieve non-destructive transient membrane permeation and transport of different macromolecules ; This work was partially supported by the Spanish Agencia Estatal de Investigación (AEI) [SAF2017-89890-R], the Xunta de Galicia (ED431C 2017/25, 2016-AD031 and Centro Singular de Investigación de Galicia accreditation 2019–2022, ED431G 2019/03), the European Union (European Regional Development Fund – ERDF) and the ISCIII (RD16/0008/003). M. P. thanks the Xunta de Galicia (ED481A-2017/142), and M. J. and G. S. thank MINECO for their F. P. I. fellowships (BES-2015-071779; PRE2018-085973). R. G.-F. thanks a RyC (RYC-2016-20335), MINECO (RTI2018-098795-A-I00) and Xunta de Galicia (ED431F 2020/05). J. M. received a RyC (RYC-2013-13784), an ERC-Stg (DYNAP-677786), a HFSP (RGY0066/2017) and ISCIII (COV20/00297). All calculations were carried out at Centro de Supercomputación de Galicia (CESGA) ; SI