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Preparation of monoclonal antibodies for monitoring radiation effects

Research Project

Project/Area Number 07557237
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field Radiation science
Research InstitutionUniversity of Tokyo

Principal Investigator

SUZUKI Norio  University of Tokyo, Faculty of Medicine, Professor, 医学部, 教授 (10010050)

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)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsMOLT-4 / X-rays / Radiation-induced proteins / p41 / Anti-p41 antibody / Cell death / Predictive assay / X線誘導蛋白p41 / p41抗体 / Set遺伝子 / MOLT4白血病細胞 / 放射線誘導蛋白 / 抗ペプチド抗体 / SET
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.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Morimatsu,A.,Suzuki,N.,et.al: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia" J.Radiat.Res.37. 1-11 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto,Y.,Suzuki,N.,et al: "DNA-dependent protein kinase activity of ataxia telangiectasia-and X-linked agammagulobulinemia-derived cells." J.Radiat.Res.37. 344-344 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto,Y.,Suzuki,N.,et al.: "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). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Morimatsu,A.,Suzuki,N.,et al: "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, 482 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Morimatsu, A., Suzuki, N., Hirano, K., Matsumoto, Y., and Sakai, K.: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia." J.Radiat. Res.37. 1-11 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto, Y,Naritaka, S., Sakai, K., and Suzuki, N.: "DNA-dependent protein kinase activity of ataxia telangiectasia-and X-linked agammaglobulinemia-derived cells." J.Radiat. Res.37. 344 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [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
    • Related Report
      1996 Final Research Report Summary
  • [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
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto,Y.et al.: "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). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Morimatsu,A.et al.: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia." J.Radiat.Res.37. 1-11 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sakata, K.: "Further application and evaluation of critical cell number and modifying factors in rario-curability of multicellular spheroids." J. Radiat. Res.36(1). 17-30 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Morimatsu, A.: "Identifibation and characterization of a protein appeared after X-irradiation in human T cell leukemia." J. Radiat. Res.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Itoh, M.: "Enhancement of heat sensitivity in FSA1233 tumor by H7" Radiation Reseach. Vol. 1. 399-399 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sakai, K.: "Modification of heat sensitivity of chiness hamster V79 cells by an inhibitor of protein kinase C" Radiation Reseach. Vol. 1. 399-399 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hirano, K.: "MUCl mucin as a radiation-inducible marker in colon carcinoma cells and its enhancement by TNF β" Radiation Reseach. Vol. 1. 226-226 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takeuchi, K.: "cDNA cloning of mouse VLA-3 αsubunit" J. Cellular Biochemistry. 57. 371-377 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 鈴木 紀夫: "癌の放射線療法'95" 大川編(T. Okawa)篠原出版, 989 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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