Ref ID : 4312
Gerene M. Denning and Alice B. Fulton; Purification and Characterization of Clathrin-Coated Vesicles from Chlamydomonas. J.Protozool. 36(4):334-340, 1989
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Clathrin-coated vesicles, identified by negative staining with uranyl acetate, were purified from Chlamydomonas reinhardtii. Isolated coated vesicles had diameters ranging from 70 to 140 nm (mean diameter+/-SD of 95+/-17 nm, n=300). These vesicles were markedly heterogeneous in both density and surface charge, as indicated by equilibrium density sedimentation and elution from anion-excharge columns. Highly-purified coated-vesicle fractions contained 2 major polypeptides, identified as the clathrin heavy chain (185 kDa) and the clathrin light chain (40 kDa). Chlamydomonas clathrin heavy chain cross-reacts weakly with an antibody against bovine brain calthrin heavy chain. Coat stability in several buffers was compared to that of bovine brain coated vesicles, stability was similar, except for a greater stability of Chlamydomonas coated vesicles in 0.5M Tris at pH 7.0.