HSV-1, which belongs to the Herpesviridae family, is responsible for different human infections. It is mainly involved in painful, cutaneous infections and in some cases can cause serious disorders such as blindness and encephalitis[1] . Fusion of host membranes with the viral envelope is necessary for the HSV-1 infection mechanism. Penetration is mediated by a group of glycoproteins conserved in all Herpesvirinae subfamilies, such as the glycoproteins B (gB), H (gH), L (gL) and D (gD). The entry process starts with the virus binding to the cell receptor and is followed by fusion between the viral envelope and a cellular membrane mediated by gB and gH/gL via conformational changes[2,3] . The purpose of this work is the evaluation of antiviral activity of several synthetic peptides derived from gH and gL sequences. In detail, we are interested to understand their specific portion involved in membrane interactions and therefore in triggering of the fusion mechanism. Therefore, we have prepared and screened two libraries of overlapping peptides homologous to the ectodomains of the two viral glycoproteins. The peptides were synthesized using the standard solid-phase Fmoc method on a an automatic synthesizer SYRO multisyntec. HPLC-MS was applied for peptides purification. We obtained 80% yields of synthesis and over than 95% of purification. Several assays of viral entry inhibitions have been performed. Between 148 peptides derived from gH, 24 of these have reached 50% of inhibition in a co-treatment assay at 150 µM as final concentration. Interestingly, they are mainly located in the gH ectodomain carboxy-terminal region, suggesting that this portion is essential for the entry mechanism. In the same time none of the gL peptides has shown a clear inhibition effect, supporting its secondary role in the membrane fusion mechanism. Our data support direct involvement of the described domains in membrane fusion, therefore those results are of relevance for the potential development of novel therapeutic compounds to prevent HSV- 1 infections.
Regulation of HSV-1 infectivity by synthetic peptides mimicking Herpes Glicoproteins / Palomba, L.; Zannella, C.; Folliero, V.; Falanga, Annarita; Martora, F.; Galdiero, M.; Vitiello, M.; Perillo, E.; Galdiero, Stefania; Morelli, Giancarlo; Galdiero, M.. - 1:(2016). (Intervento presentato al convegno 15th Naples Workshop on Bioactive Peptides tenutosi a Napoli nel 23-25 Giugno 2016).
Regulation of HSV-1 infectivity by synthetic peptides mimicking Herpes Glicoproteins
FALANGA, ANNARITA;Vitiello, M.;GALDIERO, STEFANIA;MORELLI, GIANCARLO;
2016
Abstract
HSV-1, which belongs to the Herpesviridae family, is responsible for different human infections. It is mainly involved in painful, cutaneous infections and in some cases can cause serious disorders such as blindness and encephalitis[1] . Fusion of host membranes with the viral envelope is necessary for the HSV-1 infection mechanism. Penetration is mediated by a group of glycoproteins conserved in all Herpesvirinae subfamilies, such as the glycoproteins B (gB), H (gH), L (gL) and D (gD). The entry process starts with the virus binding to the cell receptor and is followed by fusion between the viral envelope and a cellular membrane mediated by gB and gH/gL via conformational changes[2,3] . The purpose of this work is the evaluation of antiviral activity of several synthetic peptides derived from gH and gL sequences. In detail, we are interested to understand their specific portion involved in membrane interactions and therefore in triggering of the fusion mechanism. Therefore, we have prepared and screened two libraries of overlapping peptides homologous to the ectodomains of the two viral glycoproteins. The peptides were synthesized using the standard solid-phase Fmoc method on a an automatic synthesizer SYRO multisyntec. HPLC-MS was applied for peptides purification. We obtained 80% yields of synthesis and over than 95% of purification. Several assays of viral entry inhibitions have been performed. Between 148 peptides derived from gH, 24 of these have reached 50% of inhibition in a co-treatment assay at 150 µM as final concentration. Interestingly, they are mainly located in the gH ectodomain carboxy-terminal region, suggesting that this portion is essential for the entry mechanism. In the same time none of the gL peptides has shown a clear inhibition effect, supporting its secondary role in the membrane fusion mechanism. Our data support direct involvement of the described domains in membrane fusion, therefore those results are of relevance for the potential development of novel therapeutic compounds to prevent HSV- 1 infections.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
