Improved immunodetection of human papillomavirus E7.
Human papillomavirus E7 (HPV E7) is a viral oncoprotein that plays an important role in cervical carcinogenesis through binding with retinoblastoma protein (Rb). Inactivation of Rb by E7 is necessary but not sufficient for cellular transformation, suggesting other protein-protein interactions are required for E7-mediated cellular transformation aside from the interaction with Rb. However, studies on the oncogenic function of HPV E7 have been limited by its poor immunoreactivity. In this report, we show that the fixation of purified recombinant HPV E7 on blotted nitrocellulose membrane with glutaldehyde markedly enhanced the immunoreactivity of HPV E7 protein. Using HeLa and Caski cell lines which are infected with HPV 18 and HPV 16, respectively, we demonstrated that native HPV E7 proteins also could be detected by this method. These results therefore can provide the experimental conditions for detection of HPV E7 proteins with greater sensitivity and may help to analyze E7 functions.
Jeon
Ju-Hong
JH
Department of Biochemistry and Molecular Biology, Aging Apoptosis Research Center, Seoul National University College of Medicine, Seoul 110-799, Korea.
Cho
Sung-Yup
SY
Kim
Chai-Wan
CW
Shin
Dong-Myung
DM
Kwon
Joon-Cheol
JC
Choi
Kyung-Ho
KH
Kim
In-Gyu
IG
D002457:Cell Extracts / Q000737:chemistry / Q000276:immunology; D002460:Cell Line; D004268:DNA-Binding Proteins; D006801:Humans; D007120:Immunochemistry / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000032:analysis* / Q000276:immunology; D027383:Papillomaviridae / Q000737:chemistry / Q000276:immunology*; D050725:Papillomavirus E7 Proteins
D002457:Cell Extracts; D004268:DNA-Binding Proteins; C052633:E7 protein, Human papillomavirus type 18; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16