1996 Fiscal Year Final Research Report Summary
Preparation of monoclonal antibodies for monitoring radiation effects
| Project/Area Number |
07557237
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Radiation science
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| Research Institution | University of Tokyo |
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Principal Investigator |
SUZUKI Norio University of Tokyo, Faculty of Medicine, Professor, 医学部, 教授 (10010050)
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| Co-Investigator(Kenkyū-buntansha) |
HIRANO Kazuya University of Tokyo, Faculty of Medicine, Research Associate, 医学部, 助手 (80251221)
ITOH Masamitsu University of Tokyo, Faculty of Medicine, Research Associate, 医学部, 助手 (80176362)
SAKAI Kazuo University of Tokyo, Faculty of Medicine, Lecturer, 医学部, 講師 (40153837)
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| Project Period (FY) |
1995 – 1996
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| Keywords | MOLT-4 / X-rays / Radiation-induced proteins / p41 / Anti-p41 antibody / Cell death / Predictive assay |
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| Research Abstract |
The aims of the project were (1) to characterize the appearance of a protein after X-irradiation in radiosensitive human T cell leukemia MOLT-4 cells, and (2) to prepare monoclonal antibodies against the protein.
By two-dimensional electrophoresis and silver staining, a new spot was detected after irradiation. The location was 41 kDa in the mass and 4.0 of pI.The protein (p41) was identified from its partial sequences as a set gene product, whose gene translocation was previously found in a case of acute undifferentiated leukemia. Polyclonal rabbit antibodies against p41 were prepared ; the antibodies detected p41 and a 42kDa protein (p42). p42 was present also in non-irradiated control cells. Photoimage analyzes of the silver stained gels and Western blots indicated that appearance of p41 was time dependent, starting 4 hrs after irradiation and reaching a plateau by 10 hrs, and dose dependent between 1 and 10 Gy.
By the following evidences, p41 was suggested to be derived from p42 through post-translational modification ; (1) p41 appeared in the presence of cycloheximide, a protein synthesis inhibitor, (2) radioactivity was not found in p41 after post-irradiation labeling with ^<35>S-amino acids, and (3) in cells which had been pre-labeled with ^<35>S-amino acids, the radioactivity was found in p41 and p42 after irradiation. Suppression of p41 appearance after irradiation by vanadate, an inhibitor of protein phosphatase, suggested that phosphorylation/dephosphorylation would be involved in the post-translational modification.
The anti-p41 polyclonal antibodies were prepared and have been successfully utilized, as described above, for the analyzes of p41/p42. The preparation of monoclonal antibodies against p41 and p42 has been under way.
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[Publications] Matsumoto, Y., Suzuki, N., Sakai, K., Hirano, K., Morimatsu, A., and Murofushi, H.: "A possible mechanism for hyperthermic radiosensitization mediated through hyperthermic lability of Ku subunits in DNA-dependent protein kinase." Biochemical and Biophysical Research Communications. (in press).
Description
「研究成果報告書概要(欧文)」より
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[Publications] Morimatsu, A., Hirano, K., Sakai, K., Matsumoto, Y., and Suzuki, N.: Identification and characterization of an X-ray induced protein of human T cell leukemia.In "Radiation Research 1895-1995" vol. 1, Hagen, U.et al. eds, 226 (1995)
Description
「研究成果報告書概要(欧文)」より
