PMID: 1730011Jan 10, 1992

Isolation of the membranes from secretory organelles (trichocysts) of Paramecium tetraurelia

Biochimica Et Biophysica Acta
R Glas-AlbrechtH Plattner

Abstract

We present for the first time a method for isolation of the membranes of extrusive organelles (trichocysts) from sterile culture of different strains of Paramecium tetraurelia. First, trichocysts are isolated according to a new method (Glas-Albrecht, R. and Plattner, H. (1990) Eur. J. Cell Biol. 53, 164-172) with high purity and yield. Then the organelles are subjected to osmotic swelling. Since trichocysts then easily 'decondense' and entangle membranes, these cannot be isolated directly by centrifugation, but only by passage through a filter and subsequent centrifugation. Purity of membrane fractions is analysed by electron microscopy and SDS-PAGE, combined with silver staining or, after biotinylation, by avidin-peroxidase labelling. Molecular masses resolved in our gels are in a range from less than or equal to 15 to greater than or equal to 105 kDa. Main bands obtained with nd9-28 degrees C trichocyst membranes (most bands also being common to wild type trichocysts) are of about 16.5, 19-21, 27-29, 33-34, 44-45 (strong), 47-48 (strong), 57, 61, 65 (strong), 68-71, 75, 81, 94-95 (strong), 104 and greater than or equal to 110 kDa, from a total of approx. 23 bands resolved. There is no remarkable occurrence of dominant protein...Continue Reading

References

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Oct 19, 1990·European Journal of Protistology·R Glas-AlbrechtH Plattner

Related Concepts

Biodermatin
SDS-PAGE
Peroxidase-Antiperoxidase Complex Technique
Intracellular Membranes
Electron Microscopy
Phospholipids
Western Blot
Silver Stain Method
Paramecium tetraurelia

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