CALCIUM-DEPENDENT SECRETORY VESICLE-BINDING AND LIPID-BINDING PROTEINS OF SACCHAROMYCES-CEREVISIAE

Yeast (Saccharomyces cerevisiae) cytosol was examined for the presence of calcium-dependent membrane- or lipid-binding proteins that might play fundamental roles in membrane-associated phenomena in stimulated cells. A complex group of proteins was isolated from late log phase cultures of yeast strain YP3 on the basis of calcium-dependent association with yeast secretory vesicles isolated from the temperature-sensitive sec6-4 secretory mutant. The masses of the major proteins in this group were 32, 35, 47, 51, 55, 60 and 120 kDa. A similar group of proteins was isolated by calcium-dependent association with bovine brain lipids enriched in the predominant acidic phospholipids of the yeast secretory vesicles. The 47 kDa protein was highly purified when commercial yeast cake was used as the source of yeast cytosol. The 32 kDa and 60 kDa proteins were demonstrated to reassociate with lipids at calcium concentrations of 100-mu-M or higher, while no association was promoted by 2 mM-magnesium. The 47 kDa protein could be removed from lipids by reducing the calcium concentration to between 1 and 32-mu-M. The sequences of peptides isolated from digests of several of these proteins indicate that they are novel proteins but are insufficient to judge the possible homology of these proteins with mammalian membrane-binding proteins. The sequence data may be adequate to permit isolation and modification of the corresponding genes in order to assess the possible function of this class of proteins in stimulated cells.