Fractionation of Thylakoid Membranes with the Nonionic Detergent Octyl-β-d-glucopyranoside

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Abstract
The detergent octyl-.beta.-D-glucopyranoside (30 mM in 2 mM Tris-maleate, pH 7.0) preferentially extracts complexes containing protein and chlorophylls a plus b (CP) from spinach [Spinacia oleracea cv. Longstanding Bloomsdale] leaving a residue highly enriched in CP I (P700-chlorophyll a protein). Use of the detergent results in a relatively gentle extraction since little free chlorophyll is formed and since sodium dodecyl sulfate-gel electrophoresis (on 10% acrylamide) of the extract also reveals the presence of 2 minor chlorophyll a complexes (apparent MW 47,000 and 43,000) instead of the usual single complex. The major complex preserved is CP 64, a chlorophyll a/b complex (apparent MW 64,000) which is an oligomer of another chlorophyll a/b complex, CP 27, the light-harvesting complex (apparent MW 27,000). Dissociation of each complex reveals 2 polypeptides (MW, 32,000 and 28,000) and limited proteolysis confirms that those of CP 64 have the same structure as those of CP 27. An additional chlorophyll a/b complex (apparent MW, 29,000) is clearly separable from CP 27, and differs from it and CP 64 in having a higher chlorophyll a/b ratio and a single polypeptide (MW, 29,000) which differs structurally from those of the other complexes.

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