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In Vitro Reconstitution Of Exocytosis From Sea Urchin Egg Plasma Membrane And Isolated Cortical Vesicles, Joseph H. Crabb, Paul A. Modern, Robert C. Jackson
In Vitro Reconstitution Of Exocytosis From Sea Urchin Egg Plasma Membrane And Isolated Cortical Vesicles, Joseph H. Crabb, Paul A. Modern, Robert C. Jackson
Dartmouth Scholarship
We have succeeded in reconstituting an exocytotically active egg cortex fraction by recombining purified cortical vesicles (CVs) with egg plasma membrane (PM). CVs were dislodged from a suspension of egg cortex by gentle homogenization in a dissociative buffer with a pH of 9.1, and purified by two rounds of differential centrifugation. Egg PM was prepared by shearing the cortical vesicles from a cortical lawn preparation with a jet of isotonic buffer. PM lawns produced by this procedure consist of an array of CV-free PM fragments attached via their extracellular surface to a polylysine coated glass slide. When a neutralized suspension …