Epithelial cells display separate receptors for papillomavirus VLPs and for soluble L1 capsid protein

Ying Mei Qi, Shi Wen Peng, Kylie Hengst, Magnus Evander, David S. Park, Jian Zhou, Ian H. Frazer

Research output: Contribution to journalArticle

Abstract

We examined the distribution of putative receptors for papillomavirus (PV) capsid proteins on various cell types, using either Hexahis HPV6b L1 fusion protein or synthetic HPV6b virus-like particles (VLPs). Specific, saturable binding of VLPs to CV-1 cells was demonstrated using 35S-labeled VLPs, with an average receptor number of 1 x 104/cell and a binding affinity constant (K(a)) of 4 x 107 M. VLP binding was quantitated by flow cytometry using a monoclonal antibody to the L1 capsid protein. Intense staining of epithelial and mesenchymal cells was observed. Some immature bone marrow-derived cells bound VLPs weakly, while the majority of B lymphoma cells demonstrated no binding. Binding to 12 of 16 VLP receptor positive cell lines was abolished by trypsin pretreatment of cells. Removal of cellular sialic acid or O-linked oligosaccharides separately did not affect VLP binding, which was enhanced about 25% when cells were pretreated with both neuraminidase and O-glycosidase. Culture of cells with sufficient tunicamycin to inhibit Concanavalin A binding did not diminish the binding of VLPs. Denatured L1 protein, either from VLPs or expressed from Escherichia coli as a Hexahis fusion protein, bound to a trypsin-resistant structure on a range of cell types and did not block the binding of VLPs to cells. Dual-fluorescence assay with a Burkitt lymphoma line BL72 demonstrated that Hexahis L1 protein and VLPs bind to separate cell surface molecules on BL72 cells. We conclude that the first binding of PV virus to cells is via a widely distributed membrane protein receptor(s) and that subsequent processing of particles may involve other non-trypsin-sensitive structure(s) also displayed on the cell membrane.

Original languageEnglish (US)
Pages (from-to)35-45
Number of pages11
JournalVirology
Volume216
Issue number1
DOIs
StatePublished - Feb 1 1996

ASJC Scopus subject areas

  • Virology

Fingerprint Dive into the research topics of 'Epithelial cells display separate receptors for papillomavirus VLPs and for soluble L1 capsid protein'. Together they form a unique fingerprint.

  • Cite this