1996 Fiscal Year Final Research Report Summary
Identification of protein sequence which bind to phytochrome A.
| Project/Area Number |
07839015
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 時限 |
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| Research Field |
光生物学
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| Research Institution | Yokohama City University |
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Principal Investigator |
MANABE Katsushi Yokohama City University, Faculty of Science, Professor, 理学部, 教授 (10112484)
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| Project Period (FY) |
1995 – 1996
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| Keywords | phytochrome / chromophore-protein interaction / phage library / epitope |
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| Research Abstract |
A monoclonal antibody designated Mep-1, was raised against purified phytochrome A from pea. The binding of this antibody (class IgG1) to partially degraded phytochrome (114 kDa) caused a considerable increase in the far-red peak at the stationary state after red-light-irradiation. The effect reached a plateau value when the antibody and phytochrome were present in approximately equimolar amounts. The dark transformation of Pfr to Pr of the 114-kDa phytochrome was inhibited by the addition of the antibody. However, binding of the antibody to the undegraded 121-kDa phytochrome had no effects on the spectrum of the red-light-induced photostationary state. The site at which the antibody bound to phytochrome was determined to be between amino acid residues 256 and 383 of pea phytochrome A by peptide mapping. The site of the binding was further studied using phage random peptide display system and was determined to be residues from D-354 to L-357 (DAVL). A synthetic octa-peptide SADAVLPQ (corresponding to residues from 352 to 359 of the phytochrome) indeed had strong binding activity to Mep-1. The importance of the site for the chromophore-apoprotein interaction is discussed.
Experiments to find peptide sequence which specifically bind to the physiologically active Pfr form of phytochrome are in progress using the phage random peptide display library.
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[Publications] Nakazawa, M., Moriuchi, N., Wada, N., Hirose, T., Tokutomi, S., Manabe, K.: "A monoclonal antibody affects chromophore apoprotein interactions of phytochrome A" Photochem.Photobiol.65 (in press). (1997)
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