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Title: Size Exclusion-Fast-Protein-Liquid-Chromatography Separation of the Cytochrome B6f-Complex Components from Spinach Chloroplast Membranes
Author(s): M. A. Shallan
Source: Arab Journal of Biotechnology, 1998, Vol. 1, No. 1,, 29-39

Abstract: The cytochrome b6f-complex was selectively solubilized from the thylakoid of spinach chloroplast membranes using octylglucoside detergent and purified on a sucrose density gradient in the presence of the detergent. Chromatographic separation of the complex by using size exclusion- Fast-Protein-Liquid-Chromatography (sec-FPLC) system on superose 12 column under reducing and nonreducing conditions was performed. One dimensional sodium dodecyl sulfate polyacrylamide gel-electrophoresis and staining with coomassie brilliant blue was carried out to analyze the eluted peaks containing proteins. The results presented in this work showed that the cytochrome complex may be found in an oligemeric form. A group of high molecular weight protein subuints with molecular masses ranging from approx. 37 to 95 kDa could be isolated and assumed to be in a close association with the complex in the thylakoid membrane. In addition, a binary subcomplex consisted of cytochrome b6 and the so called subunit 17 kDa was obtained and separated. The functional capacity of these higher molecular weight protein subunits as well as the observed binary subcomplex is an open question...

Addresses: Biochemistry Dept. Fac. Agriculture, Cairo Univ. Giza, Egypt...

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Country Code: EG