Critical role for cholesterol in Lassa fever virus entry identified by a novel small molecule inhibitor targeting the viral receptor LAMP1
Fig 2
Binding of LASV GP to LAMP1 is dependent on cholesterol and is inhibited by 3.3.
(A) 3.3 blocks the interaction of LASV GP with endogenous LAMP1. 293T LAMP1 KO cells expressing LASV GP-His and 293T pX459 cells were incubated with the indicated concentrations of 3.3 for 1h at 37°C before lysis. Lysates were mixed and subjected to immunoprecipitation with an anti-His antibody. The presence of LASV GP bound LAMP1 was detected by immunoblot with an anti-LAMP1 antibody. (B) LASV GP1-IgG binds to LAMP1 D1 in a cholesterol-dependent manner. Purified LAMP1 D1-His was incubated with the indicated concentrations of cholesterol or epicholesterol for 1h at 37°C before the addition of purified LASV GP1-IgG. Samples were subjected to immunoprecipitation against human IgG and bound LAMP1 D1 was detected by immunoblot with an anti-His antibody. (C) 3.3 inhibits the cholesterol-dependent binding of LASV GP1-IgG to LAMP1 D1. Purified LAMP1 D1-His was co-incubated with 5μM cholesterol and the indicated concentrations of 3.3 or 102 for 1h at 37°C before the addition of purified LASV GP1-IgG. Samples were subjected to immunoprecipitation against human IgG and bound LAMP1 D1 was detected by immunoblot with an anti-His antibody.