Immunochemical Investigation of Thylakoid Coupling Factor Protein during Photosynthetic Acclimation to Irradiance

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Abstract
Davies, E. C, Jordan, B. R., Partis, M. D. and Chow, W. S. 1987. Immunochemical investigation of thylakoid coupling factor protein during photosynthetic acclimation to irradiance.—J. exp. Bot. 38: 1517—1527 A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) has been used to determine the amount of coupling factor protein (CF1 present on thylakoid membranes isolated from lettuce leaves. Thylakoid CF1 was purified from lettuce chloroplasts and used to raise antibodies. After purification of the IgG its specificity to CF1 was examined by Ouchterlony double diffusion, Western blotting and control ELISA experiments. The specificity of the response for CF1 was confirmed and the ELISA found to give a log-linear measurement of purified CF1 concentrations within a range of 10 μg cm⊟3 to 25 ng cm ⊟3. The ELISA was then used to determine the level of CF1 protein associated with the thylakoid membrane. Comparable values were obtained for CF1 levels, either assayed directly on the thylakoid or after release from the membrane by chloroform treatment. Determination of CF1 protein content on thylakoids isolated from lettuce grown at different irradiances showed that the amount of CF1 protein per chlorophyll and chloroplast increased with increased lrradiance. In addition, the changes in CF1 protein as determined by ELISA are reflected in changes that have been found in enzymic activity under these same light environments. The implications for the photoregulation of CF1 are discussed.

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