Aggregation and dispersity of isolated chromaffin granules studied by intensity fluctuation spectroscopy

The aggregation and dispersity of isolated bovine adrenal secretory vesicles (chromaffin granules) were studied by intensity fluctuation spectroscopy. The degree of dispersity and the Z-average translational diffusion coefficients were calculated from the autocorrelation functions of the intensity fluctuations in laser light scattered from the granules in solution. Granules purified by sedimentation through 0.3 M sucrose/Ficoll/2H2O showed greater dispersity than granules purified by sedimentation through 1.6 M sucrose. By monitoring the scattered light intensity and the diffusion coefficients of the granules, many of the difficulties encountered in the interpretation of absorbance measurements were avoided. Measurements over a range of granule concentrations in sucrose solutions (10 mM HERPES, pH 7.0), indicated that aggregation of the granules occurred at concentrations above 150 microgram protein/ml. At low granule concentrations (15-30 microgram protein/ml) Ca2+-induced aggregation was detected at a threshold of 2-10 mM calcium.