Photosynthetica 1997, 33(13):447-464 | DOI: 10.1023/A:1006876420980

Flow cytometric characteristics of subchloroplast particles prepared by the action of various detergents

N. Wilhelmová1, S. Vanìk2, J. Wilhelm3
1 Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Praha 6, Czech Republic
2 Department of Clinical Haematology of the Faculty Hospital of Charles University, Praha 2, Czech Republic
3 2nd Medical School, Charles University, Praha 5, Czech Republic

Agranal thylakoid membranes from leaves of Phaseolus vulgaris L. were fragmented using seven distinct detergents: digitonin, Triton X-100, cetylpyridinium chloride, sodium dodecylsulfate, and Zwittergents 3-12, 3-14, and 3-16, differing in chemical composition and/or electric charges. Subchloroplast particles separated on a Percoll gradient were examined by flow cytometry to determine their size and shape. Vesicle size was also determined by a haematological analyzer, which produced comparable results. Individual green bands consisted of vesicles of fairly wide size distribution. Simple direct proportionality between the particle density and their size was not observed in any case, nevertheless, bigger particles were more abundant in fractions of higher density. Some vesicles had even a larger size than the original thylakoids. This might reflect a specific action of the detergents in low concentrations on agranal membranes, with incorporation of detergent molecules into vesicles. Inner structures of particles of the same size and density were not necessarily identical, but represented several populations, as was apparent from the side scatter analysis. Flow cytometric analysis can thus be used for the investigation of mechanisms of membrane fragmentation by detergents.

Additional key words: cetylpyridinium chloride; digitonin; membrane vesicles; Phaseolus vulgaris; sodium dodecylsulfate; thylakoids; Triton X-100; Zwittergents

Prepublished online: January 1, 1998; Published: September 1, 1997  Show citation

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Wilhelmová, N., Vanìk, S., & Wilhelm, J. (1997). Flow cytometric characteristics of subchloroplast particles prepared by the action of various detergents. Photosynthetica, 34(3), 447-464. doi: 10.1023/A:1006876420980
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