Abstract
Upon protoolytle treatment of thylakoid membranes, extrinsic proteins are selectively degraded. The remaining resistant proteins have been analyzed by SDS-PAGE. In the thylakoids of Chlamydomonas reinhardii, six polypeptides or protein fragments of 20 kD or higher are resistant to proteolysis. These intrinsic proteins have been identified as: the apoproteins of the chlorophyll-protein complexes CP I and LHCP; a polypeptide whose presence is related to the chlorophyll b content of the cells; and a portion of a chloroplast-made 34 kD polypeptide. Furthermore, after proteolytic treatment of the membranes, the LHCP complex can be resolved into two subcomplexes, apparently differing in their polypeptide composition.
Zusammenfassung
Durch proteolytische Behandlung von Thylakoidmembranen worden die extrinsischen Proteinanteile selektiv abgebaut. Die resistenten, in der Membran verbleibenden Polypeptide können mit SDS-PAGE analysiert werden. In Thylakoiden von Chlamydomonas reinhardii finden sich sechs resistente Polypeptide mit Molekulargewichten über 20'000. Durch quantitative Bestimmung während der Proteolyse und durch limitierte Fragmentierung nach Cleveland wurden die resistenten Polypeptide identifiziert als: (a) Apoprotein von CP 1, (b) Apoproteine des LHCP-Komplexes, (c) Teil des chloroplastidial synthetisierten 34 kD-Proteins und (d) Polypeptid, welches möglicherweise mit dem Chlorophyll b-Gehalt in Verbindung steht. Im weiteren führte die proteolytische Behandlung der Thylakoide zu einer Auftrennung des LHCP-Komplexes in zwei Subkomplexe mit unterschiedlicher Proteinzusammensetzung.
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Abbreviations
- CP 1:
-
pigment-protein complex with slower mobility in SDS electrophoresis, corresponding to the P700 chlorophyll a protein complex
- DCMU:
-
3-(3,4-dichlorophenyl)-1,1-dimethyl urea
- EDTA:
-
ethylene-diaminotetraacetic acid
- kD:
-
kilo Dalton
- LHCP:
-
pigment-protein complex with intermediate mobility in SDS electrophoresis, corresponding to the light-harvesting chlorophyll a/b protein complex
- SDS:
-
sodiumdodecyl sulfate
- SDS-PAGE:
-
electrophoresis in SDS-containing polyacrylamide gels
- Tris:
-
tris (hydroxymethyl) aminomethane
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Michel, H.P., Schneider, E., Tellenbach, M. et al. Intrinsic membrane proteins of the thylakoids of Chlamydomonas reinhardii . Photosynth Res 2, 203–212 (1981). https://doi.org/10.1007/BF00032359
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DOI: https://doi.org/10.1007/BF00032359