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Carboxy-fluorescein...
Carboxy-fluorescein diacetate, succinimidyl ester labeled papillomavirus virus-like particles fluoresce after internalization and interact with heparan sulfate for binding and entry
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- Drobni, Peter (author)
- Umeå universitet,Virologi
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- Mistry, Nitesh (author)
- Umeå universitet,Virologi
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McMillan, Nigel (author)
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- Evander, Magnus (author)
- Umeå universitet,Virologi
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(creator_code:org_t)
- 2003
- 2003
- English.
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In: Virology. - 0042-6822 .- 1096-0341. ; 310:1, s. 163-172
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- Human papillomaviruses (HPVs) infect epithelial cells and are associated with genital carcinoma. Most epithelial cell lines express cell-surface glycosaminoglycans (GAGs) usually found attached to the protein core of proteoglycans. Our aim was to study how GAGs influenced HPV entry. Using a human keratinocyte cell line (HaCaT), preincubation of HPV virus-like particles (VLPs) with GAGs showed a dose-dependent inhibition of binding. The IC(50) (50% inhibition) was only 0.5 microg/ml for heparin, 1 microg/ml for dextran sulfate, and 5-10 microg/ml for heparan sulfate from mucosal origin. Mutated chinese hamster ovary (CHO) cell lines lacking heparan sulfate or all GAGs were unable to bind HPV VLPs. Here we also report a method to study internalization by using VLPs labeled with carboxy-fluorescein diacetate, succinimidyl ester, a fluorochrome that is only activated after cell entry. Pretreatment of labeled HPV VLPs with heparin inhibited uptake, suggesting a primary interaction between HPV and cell-surface heparan sulfate.
Keyword
- Papillomavirus; CFDA SE; Heparan sulfate; Uptake
- Receptor
Publication and Content Type
- ref (subject category)
- art (subject category)
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