Release of hepatitis B virions is positively regulated by glucose‐regulated protein 78 through direct interaction with preS1

In this study, we investigated the mechanism of hepatitis B virus (HBV)‐enveloped particle release. Specifically, we used preS1 as a bait protein to screen host proteins using mass spectroscopy, with the results of immunofluorescence, western blot, co‐immunoprecipitation, isothermal titration calorimetry, and pull‐down assays identifying glucose‐regulated protein (GRP)78 as a specific target for preS1 binding. We employed transcriptome sequencing, enzyme‐linked immunosorbent assays, and particle gel assays to investigate the mechanism of GRP78‐mediated positive regulation of HBV‐enveloped particle release. Additionally, we performed phage‐display, surface plasmon resonance, and molecular‐docking assays to assess peptides inhibiting enveloped‐particle release. We found that HBV upregulated GRP78 expression in liver cell lines and the serum of patients with chronic hepatitis B. Furthermore, GRP78 promoted the release of HBV‐enveloped particles in vitro and in vivo within an HBV transgenic mouse model. Moreover, we identified interactions of preS1 peptides with GRP78 via hydrogen bonding and hydrophobic interactions, which effectively inhibited its interaction with HBV‐enveloped particles and their subsequent release. These findings provide novel insights regarding HBV virion release, and demonstrated that GRP78 interacted with preS1 to positively regulate the release of HBV‐enveloped particles, suggesting GRP78 as a potential therapeutic target for inhibiting HBV infection.