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Journal of General Virology | 1994

Functional oligomerization of purified human papillomavirus types 16 and 6b E7 proteins expressed in Escherichia coli

Masanobu Chinami; Shigeo Sasaki; Naoki Hachiya; Kentaro Yuge; Takeo Ohsugi; Hiroshi Maeda; Masahisa Shingu

Purified non-fused soluble human papillomavirus type 16 and 6b E7 proteins expressed in Escherichia coli were found to form oligomers. For both proteins, several degrees of oligomerization were demonstrated by gel filtration, dynamic laser light scattering and scanning electron microscopy. Oligomerization was dependent on the concentration of E7 protein. Oligomerized E7 proteins were able to bind the retinoblastoma gene product pRB and stimulated DNA synthesis when introduced into cells.


Protein Expression and Purification | 1991

Refolding and purification of human papillomavirus type 16 E7-IacZ fusion protein expressed in Escherichia coli

Masanobu Chinami; Kentaro Yuge; Keiichi Kawano; Masahisa Shingu

Human papillomavirus (HPV) type 16 E7-lacZ fusion protein was produced in Escherichia coli, extracted as inclusion bodies, refolded with reducing reagents, and subjected to gel filtration. The refolded protein was purified by ion-exchange column chromatography, resulting in a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1H nuclear magnetic resonance spectral changes were observed in the high field methyl region in the presence of Zn2+ ion, suggesting that the refolded form of the fusion protein is possibly renaturated into the putative zinc finger motif (C. Edmond and K. H. Vousden, 1989, J. Virol. 63, 2650-2656) and supporting the data of J. A. Rawls, R. Pusztai, and M. Green (1990, J. Virol. 64, 6121-6129) on zinc binding to E7 protein using radioisotopically labeled zinc ion.


Journal of Virological Methods | 1996

Nucleic acid binding by zinc finger-like motif of human papillomavirus type 16 E7 oncoprotein

Masanobu Chinami; Masahiro Inoue; Kenji Masunaga; Toshihide Fukuma; Masahisa Shingu; Tetsuya Toyoda

Human papillomavirus (HPV) types 16 and 6b E7 proteins and their chimeric or mutant proteins were analyzed for oligonucleotide-binding activity by surface plasmon resonance-based biomolecular interaction analysis. The results indicated that type 16 E7 protein has stronger nucleic acid-binding activity than that of type 6b E7 protein. In addition, the results also indicated that the zinc finger-like motif in the C-terminal region of the type 16 E7 protein plays an important role in this activity.


Archives of Dermatological Research | 1990

Human papillomavirus DNA from warts for typing by endonuclease restriction patterns: purification by alkaline plasmid methods

Masanobu Chinami; Eiko Tanikawa; Hiroshi Hachisuka; Yoichiro Sasai; Masahisa Shingu

SummaryThe alkaline plasmid DNA extraction method of Birnboim and Doly was applied for the isolation of human papillomavirus (HPV) from warts. Tissue from common and plantar warts was digested with proteinase K, and the extrachromosomal circular covalently-closed form of HPV-DNA was rapidly extracted by alkaline sodium dodecyl sulphate and phenol-chloroform treatment. Recovery of HPV-DNA from the tissue was sufficient for determination of endonuclease restriction patterns by agarose gel electrophoresis.


Autoimmunity | 1993

Significance of anti-eye muscle antibody in patients with thyroid-associated ophthalmopathy by quantitative western blot

Yuji Hiromatsu; Masayuki Sato; Kiyoko Tanaka; Shingo Shoji; Kyohei Nonaka; Masanobu Chinami; Hiroshi Fukazawa


Archives of Biochemistry and Biophysics | 1989

Hydrogen-1 nuclear magnetic resonance studies of staphylococcal nuclease variant H124L: pH dependence of histidines and tyrosines

Masanobu Chinami; Masahisa Shingu


The Kurume Medical Journal | 1993

Experimental Transmission of HTLV-I into C3H/HeJ Mice

Takeo Ohsugi; Masanobu Chinami; Kentaro Yuge; Naoki Hachiya; Masahisa Shingu


The Kurume Medical Journal | 1994

Transformation Assays of the Cell Mitotic and Proliferation Activities of Purified Oncogene Products

Kentaro Yuge; Masanobu Chinami; Masahisa Shingu


The Kurume Medical Journal | 1992

Construction of an Expression Vector for Human Papillomavirus Type 16 E7-Glutathion S Transferase Fusion Protein

Masanobu Chinami; Masahisa Shingu


The Kurume Medical Journal | 1986

Poliovirus Resistant Cells Derived from HeLa Cells

Masanobu Chinami; Eiji Nakamura; Shinichi Kakisako; Bing Xu; Masahisa Shingu

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