Title
A new cryptic host defense peptide identified in human 11-hydroxysteroid dehydrogenase-1 β-like: from in silico identification to experimental evidence
Author
A. Bosso, L. Pirone, R. Gaglione, K. Pane, A. Del Gatto, L. Zaccaro, S. Di Gaetano, D. Diana, R. Fattorusso, E. Pedone, V. Cafaro, H.P. Haagsman, A. van Dijk, M.R. Scheenstra, A. Zanfardino, O. Crescenzi, A. Arciello, M. Varcamonti, E.J.A. Veldhuizen, A. Di Donato, E. Notomista, E. Pizzo
Year
2017
Journal
Biochimica et Biophysica Acta (BBA) - General Subjects
Abstract
Host defence peptides (HDPs) are evolutionarily conserved components of innate immunity. Human HDPs, produced by a variety of immune cells of hematopoietic and epithelial origin, are generally grouped into two families: beta structured defensins and variably-structured cathelicidins. We report the characterization of a very promising cryptic human HDP, here called GVF27, identified in 11-hydroxysteroid dehydrogenase-1 β-like protein. Conformational analysis of GVF27 and its propensity to bind endotoxins were performed by NMR, Circular Dichroism, Fluorescence and Dynamic Light Scattering experiments. Crystal violet and WST-1 assays, ATP leakage measurement and colony counting procedures were used to investigate antimicrobial, anti-biofilm, cytotoxicity and hemolytic activities. Anti-inflammatory properties were evaluated by ELISA. GVF27 possesses significant antibacterial properties on planktonic cells and sessile bacteria forming biofilm, as well as promising dose dependent abilities to inhibit attachment or eradicate existing mature biofilm. It is unstructured in aqueous buffer, whereas it tends to assume a helical conformation in mimic membrane environments as well as it is able to bind lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Notably it is not toxic towards human and murine cell lines and triggers a significant innate immune response by attenuating expression levels of pro-inflammatory interleukins and release of nitric oxide in LPS induced macrophages. Human GVF27 may offer significant advantages as leads for the design of human-specific therapeutics.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Chemical stability, Vesicle interactions, Biochemistry