| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
The process of outer segment disk formation occurs throughout the life of an organism, providing a mechanism for continual renewal of outer segment components. On a disk membrane with dynamic structure, a lot of proteins related to phototransduction, such as rhodopsin, exist and the transduction cascade works when the segment has received light. Since the cascade reaction progresses very quickly, formation of micromachinery that accumulates the proteins can be predicted. Immunocytochemistry revealed that tubulin alpha, beta, gamma and tau were co-localized on the disk membrane. To identify the tubulin specific binding proteins in outer segments, we demonstrated immunoprecipitation and mass spectrometry. Some proteins were identified by mass spectrometry. The tau and an annexin II were co-precipitated with tubulin beta and gamma. These results suggest that the tubulin plays the role of the scaffold of a phototransduction micromachinary. Since the bovine retina stopped obtaining for the BSE of the bovine, we changed material into the frog retina. We prepared frog ROS fraction and analyzed the fraction by 2D-PAGE and mass spectrometry. Thirty five proteins had been identified when 136-gel spots were analyzed. Forteen proteins were rhodopsin, The proteins of 14 originated in outer segment and contained rhodopsin, phosphodiesterase alpha and beta, transducin alpha and beta, arrestin and s-modulin. The proteins of 18 originated in inner segment and contained HSC70,enolase, GTP-rho binding protein, neuronal intermediate filament and ATP synthase. The actin, and a tubulin alpha and beta were common to both.
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