gH625 is a viral derived peptide for effective delivery of intrinsically disordered proteins

Giovanni Smaldone,^1,2 Annarita Falanga,³ Domenica Capasso,⁴ Daniela Guarnieri,^5,6 Stefania Correale,^1,7 Massimiliano Galdiero,⁸ Paolo A Netti,⁴ Massimo Zollo,⁹ Stefania Galdiero,^1,2 Sonia Di Gaetano,¹ Emilia Pedone<sup>1

1</sup>Institute of Biostructures and Bioimaging, National Research Council, Naples, Italy; ²Department of Pharmacy and Interuniversity Research Center on Bioactive Peptides, Federico II University of Naples, Naples, Italy; ³Molecular Diagnostics and Pharmaceuticals Scarl, Naples, Italy; ⁴Special Center for Biotechnology, Federico II University of Naples, Naples, Italy; ⁵Center for Advanced Biomaterials for Health Care, Interdisciplinary Research Centre on Biomaterials, Italian Institute of Technology, Naples, Italy; ⁶Interdisciplinary Research Centre on Biomaterials, Federico II University of Naples, Naples, Italy; ⁷Kedrion S.p.A, Sant'antimo, Naples, Italy; ⁸Department of Experimental Medicine, Federico II University of Naples, Naples, Italy; ⁹CEINGE – Advanced Biotechnology Scarl, Naples, Italy

Abstract: A genetically modified recombinant gH625-c-prune was prepared through conjugation of c-prune with gH625, a peptide encompassing 625–644 residues of the glycoprotein H of herpes simplex virus 1, which has been proved to possess the ability to carry cargo molecules across cell membranes. C-prune is the C-terminal domain of h-prune, overexpressed in breast, colorectal, and gastric cancers, interacting with multiple partners, and representing an ideal target for inhibition of cancer development. Its C-terminal domain results in an intrinsically disordered domain (IDD), and the peculiar properties of gH625 render it an optimal candidate to act as a carrier for this net negatively charged molecule by comparison with the positively charged TAT. A characterization of the recombinant gH625-c-prune fusion protein was conducted by biochemical, cellular biology and confocal microscopy means in comparison with TAT-c-prune. The results showed that the gH625-c-prune exhibited the ability to cross biomembranes, opening a new scenario on the use of gH625 as a novel multifunctional carrier.

Keywords: delivery, IDP