Detection of E5 oncoprotein in human papillomavirus type 16-positive cervical scrapes using antibodies raised to synthetic peptides.
Polyclonal antibodies were raised to partial and full-length synthetic peptides of human papillomavirus type 16 (HPV-16) E5. Antisera specificity for HPV-16 E5 was demonstrated by their ability to recognize not only their peptide immunogens but also full-length peptide and a glutathione S-transferase-E5 fusion protein. The most reactive antiserum, PE-6, raised to a full-length peptide, was used in Western blot analysis to identify HPV-16 E5 protein from exfoliated cervical cells. A strong, single band at approximately 20K was detected in two of six HPV-16-positive samples from women with a history of low-grade cervical intraepithelial neoplasia. The apparent M(r) by SDS-PAGE suggests that HPV-16 E5 forms homodimers in vivo, but not through cysteine linkage.
Kell
B
B
Richard Dimbleby Laboratory of Cancer Virology, Rayne Institute, United Medical School, London, U.K.
Jewers
R J
RJ
Cason
J
J
Pakarian
F
F
Kaye
J N
JN
Best
J M
JM
D000914:Antibodies, Viral; D015153:Blotting, Western; D002584:Cervix Uteri / Q000821:virology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D005982:Glutathione Transferase / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D027383:Papillomaviridae / Q000276:immunology / Q000302:isolation & purification*; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology; D012097:Repressor Proteins; D014626:Vaginal Smears
D000914:Antibodies, Viral; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D010455:Peptides; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D005982:Glutathione Transferase