Herpes simplex virus type 1 (HSV-1) is a member of the α-herpesvirinae subfamily that infects mucocutaneous tissues in a latent way to cause cold sores and, under certain circumstances, may cause encephalitis [1]. It enters cells through fusion of the viral envelope with a cellular membrane involving multiple interactions between viral glycoproteins and cellular receptors. The envelope glycoprotein B, D, H and L are all essential for the entry process [2]. In particular, gB and gH/gL interact with each other concomitantly with fusion and this interaction is triggered by binding of gD to its cellular receptor. The glycoprotein B is the key fusion protein [2]. Its crystal structure , composed by a trimer, shows that each protomer is divided into five distinct domains; the domain III contains a trimeric coiled coil and a long nonhelical C-terminal arm packing against the coiled coil in an antiparallel mode [3]. Previous results from our laboratory have shown that a peptide comprising the N-terminus of the helical sequence (residues from 500 to 523, named gBhN) is an active inhibitor of HSV-1 infection prior to virus penetration into cells [3]. The aim of this work is to improve the antiviral activity of the peptide gBhN through binding it to a cholesterol tail[4][5]. Solid phase-9-fluorenyl-methoxycarbonyl (Fmoc) on an automatic synthetizer SYRO multisyntech standard method for peptide synthesis has been used. Then the peptides has been purified with HPLC, characterized with HPLC/MS and bound to cholesterol. A citotoxicity assay has been realized, while a co-treatment and a cell pre-treatment assays have been performed to test antiviral activity of the complex. The compounds have not shown any toxicity on eukaryotic cells, even at concentrations higher than the ones used in antiviral testing. Moreover, we have obtained an improvement in the antiviral effect of the complex gBhN/cholesterol compared to the peptide alone. In conclusions, our results provide some new spotlights in HSV-1 entry mechanism but further studies are in progress to deepen the mechanisms underlying the interaction between the peptide and cellular membrane.
Herpes Simplex Virus type 1: new antiviral strategies / Zannella, C.; Lombardi, L.; Falanga, Annarita; Galdiero, Stefania; Nocerino, A.; Folliero, V.; Dolzatelli, S.; Pessi, A.; Porotto, M.; Morelli, Giancarlo; Galdiero, M.. - (2016). (Intervento presentato al convegno 15th Naples Workshop on Bioactive Peptides nel 23-25 Giugno 2016).
Herpes Simplex Virus type 1: new antiviral strategies
FALANGA, ANNARITA;GALDIERO, STEFANIA;MORELLI, GIANCARLO;
2016
Abstract
Herpes simplex virus type 1 (HSV-1) is a member of the α-herpesvirinae subfamily that infects mucocutaneous tissues in a latent way to cause cold sores and, under certain circumstances, may cause encephalitis [1]. It enters cells through fusion of the viral envelope with a cellular membrane involving multiple interactions between viral glycoproteins and cellular receptors. The envelope glycoprotein B, D, H and L are all essential for the entry process [2]. In particular, gB and gH/gL interact with each other concomitantly with fusion and this interaction is triggered by binding of gD to its cellular receptor. The glycoprotein B is the key fusion protein [2]. Its crystal structure , composed by a trimer, shows that each protomer is divided into five distinct domains; the domain III contains a trimeric coiled coil and a long nonhelical C-terminal arm packing against the coiled coil in an antiparallel mode [3]. Previous results from our laboratory have shown that a peptide comprising the N-terminus of the helical sequence (residues from 500 to 523, named gBhN) is an active inhibitor of HSV-1 infection prior to virus penetration into cells [3]. The aim of this work is to improve the antiviral activity of the peptide gBhN through binding it to a cholesterol tail[4][5]. Solid phase-9-fluorenyl-methoxycarbonyl (Fmoc) on an automatic synthetizer SYRO multisyntech standard method for peptide synthesis has been used. Then the peptides has been purified with HPLC, characterized with HPLC/MS and bound to cholesterol. A citotoxicity assay has been realized, while a co-treatment and a cell pre-treatment assays have been performed to test antiviral activity of the complex. The compounds have not shown any toxicity on eukaryotic cells, even at concentrations higher than the ones used in antiviral testing. Moreover, we have obtained an improvement in the antiviral effect of the complex gBhN/cholesterol compared to the peptide alone. In conclusions, our results provide some new spotlights in HSV-1 entry mechanism but further studies are in progress to deepen the mechanisms underlying the interaction between the peptide and cellular membrane.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
