1887

Abstract

Baculovirus occlusion-derived virions (ODVs) are released from occlusion bodies by the alkaline environment of the insect midgut. The ODV envelope protein P74 is required for oral infectivity. A soluble form of the P74 protein, P74, was engineered as part of a chimeric protein with jellyfish green fluorescent protein (GFP). P74–GFP was overproduced by the baculovirus expression system and purified away from the wild-type P74. Brush border membrane vesicles (BBMVs) were prepared from the midguts of third-instar larvae. When P74–GFP was incubated under alkaline conditions with BBMVs, a P74–GFP product with a smaller molecular mass was produced. Immunoblots indicated that the smaller product was generated by N-terminal cleavage of P74. This cleavage was prevented by soybean trypsin inhibitor. Analysis of the peptide sequences of P74 homologues identified a conserved trypsin cleavage site that could generate the observed P74–GFP BBMV-specific cleavage product.

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2005-06-01
2020-10-23
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