Phycobilisomes from blue-green and red algae: isolation criteria and dissociation characteristics.
A general procedure for the isolation of functionally intact phycobilisomes was devised, based on modifications of previously used procedures. It has been successful with numerous species of red and blue-green algae (Anabaena variabilis, Anacystis nidulans, Agmenellum quadruplicatum, Fremyella diplosiphon, Glaucosphaera vacuolata, Griffithsia pacifica, Nemalion multifidum, Nostoc sp., Phormidium persicinum, Porphyridium cruentum, P. sordidum, P. aerugineum, Rhodosorus marinus). Isolation was carried out in 0.75 molar K-phosphate (pH 6.8 to 7.0) at 20 to 23 C on sucrose step gradients. Lower temperature (4 to 10 C) was usually unfavorable resulting in uncoupling of energy transfer and partial dissociation of the phycobilisomes, sometimes with complete loss of allophycocyanin. Intact phycobilisomes were characterized by fluorescence emission peaks of 670 to 675 nanometers at room temperature, and 678 to 685 nanometers at liquid nitrogen temperature. Uncoupling and subsequent dissociation of phycobilisomes, in lowered ionic conditions, varied with the species and the degree of dissociation but occurred preferentially between phycocyanin and allophycocyanin, or between phycocyanin and phycoerythrin.
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Radiation Biology Laboratory, Smithsonian Institution, 12441 Parklawn Drive, Rockville, Maryland 20852.
Lipschultz
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Grabowski
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Zimmerman
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