Isolation and Characterization of the Cytochrome bf Complex from Whole Thylakoids, Grana, and Stroma Lamellae Vesicles from Spinach Chloroplasts : MEMBRANES AND BIOENERGETICS

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概要

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• The cytochrome bf complex was isolated from spinach thylakoids, and also from separated grana and stroma lamellae vesicles, by a procedure involving NaBr washing, detergent treatment and centrifugation in sucrose gradients. The resulting complex from all three types of membranes, were almost completely devoid of chlorophyll and carotenoids. The complexes have kinase activity towards histone III-S and contain a 64 kDa protein claimed to be a kinase. Electrophoretic analyses indicate that the complexes are in dimeric form and composed of six poly-peptides with molecular masses of 34/33, 23, 20, 17, 12 and 4 kDa. The complexes contain two moles cytochrome b_6 per mole cytochrome f and one mole Risque FeS. The 17 kDa and 4 kDa polypeptides are the so called subunit 4 and 5 respectively. The 12 kDa protein was identified as plastocyanin by immunoblotting. Plastocyanin and the 4 kDa protein were present in the cytochrome bf complex even after a second repeated sucrose density gradient centrifugation. The sucrose gradient sedimentation pattern was different for the grana and stroma lamellae complexes. The complex from the stroma lamellae arrives at a higher density than the grana complex. Furthermore, the gradient centrifugation diagram of the stroma lamellae consists of one main peak while the diagram of the grana complex shows two peaks. There is significantly more plastocyanin and 4 kDa protein in the bf complex isolated from stroma lamellae than from grana. In addition there is a 15 kDa protein in the complex isolated from the grana vesicles. Immunoblot analysis after crosslinking indicated that the 4 kDa protein and the plastocyanin are associated in the cytochrome bf complex. The oxidoreductase activity is higher (about 50%) in the cytochrome bf complex from the grana than from the stroma lamellae fraction. We suggest that a difference in composition of the cytochrome bf complex between the two membranes might be important in the regulation of cyclic and non cyclic electron flow.

• 日本植物生理学会の論文

著者

• Romanowska E

  Warsaw Univ. Warsaw Pol

• Romanowska Elzbieta

  Department Of Plant Physiology Botanical Institute Warsaw University

• ALBERTSSON Per-Ake

  Department of Biochemistry, University of Lund

• Albertsson Per-ake

  Department Of Biochemistry University Of Lund

• Albertsson Per-ake

  Department Of Biochemistry Chemical Center University Of Lund

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