| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1991: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
An assay system for physiological function of connectein, a constituent of the large form of ferredoxin-NADP reductase(FNR-L), was established by modifying the reconstituted NADP photoreducing system originally designed for demonstrating physiological function of FNR-L. FNR-L was highly purified by an improved method and depleted thylakoids were prepared by brief extraction of FNR-L with a detergent, CHAPS. Using these components, NADP photoreducing system was successfully reconstituted with good reproducibility. NADP photoreducing activity of thylakoids was decreased when FNR-L was removed by CHAPS treatment and the decreased activity was fully restored by rebinding of FNR-L. In contrast to FNR-L, monomer FNR, which does not associated with connection, was almost ineffective in restoring the NADPphotoreducing activity of FNR-L-depleted thylakoids. The clear defference in restoration of the NADP photoreducing activity observed between FNR with and without connection reflects functional contribution of connection to the NADP photoreducing reaction of thylakoids. Thus, physiological activity of connection could be quantified by this reconstitution experiment.
An attempt was made to analyze an amino acid composition and amino-terminal structure of connectein, since the unique function of the protein may originate from its structure. Its amino-terminal structure was identified up to the 16th residue but showed no sequence homology with any of the polypeptides sequenced so far.
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