Evidence for a specialized localization of the chloroplast ATP-synthase subunits alpha, beta, and gamma in the eyespot apparatus of Chlamydomonas reinhardtii (Chlorophyceae)

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Details zur Publikation

Autorinnen und Autoren: Kreimer G
Zeitschrift: Journal of Phycology
Verlag: Wiley-Blackwell
Jahr der Veröffentlichung: 2007
Band: 43
Heftnummer: 2
Seitenbereich: 284-294
ISSN: 0022-3646


The eyespot apparatus (EA) of Chlamydomonas reinhardtii P. A. Dang. consists of two layers of carotenoid-rich lipid globules subtended by thylakoids. The outermost globule layer is additionally associated with the chloroplast envelope membranes and the plasma membrane. In a recent proteomic approach, we identified 202 proteins from isolated EAs of C. reinhardtii via at least two peptides, including, for example, structural components, signalling-related proteins, and photosynthetic-related membrane proteins. Here, we have analyzed the proteins of the EA with regard to their topological distribution using thermolysin to find out whether the arrangement of globules and membranes provides protection mechanisms for some of them. From about 230 protein spots separated on two-dimensional gels, the majority were degraded by thermolysin. Five major protein spots were protected against the action of this protease. These proteins and some that were degradable were identified by mass spectrometry. Surprisingly, the thermolysin-resistant proteins represented the alpha and beta subunits of the soluble CF1 complex of the chloroplast ATP synthase. Degradable proteins included typical membrane proteins like LHCs, demonstrating that thermolysin is not in general sterically prevented by the EA structure from reaching membrane-associated proteins. A control experiment showed that the CF1 complex of thylakoids is efficiently degraded by thermolysin. Blue native PAGE of thermolysin-treated EAs followed by SDS-PAGE revealed that the alpha and beta subunits are present in conjunction with the gamma subunit in a thermolysin-resistant complex. These results provide strong evidence that a significant proportion of these ATP-synthase subunits have a specialized localization and function within the EA of C. reinhardtii.

FAU-Autorinnen und Autoren / FAU-Herausgeberinnen und Herausgeber

Kreimer, Georg Prof. Dr.
Professur für Zellbiologie

Zuletzt aktualisiert 2018-10-08 um 10:53

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