Figure 9.

Nonreversibility of calcium effect. Lanes 1–5: Isolated granules were treated as in Figure 7, with two additional variables. In lane 4, 10 mM EGTA was added after the calcium incubation, to reduce free Ca²⁺ to <1 μM. In lane 5, 10 mM EGTA was added before calcium. Samples were then incubated with chymotrypsin. Gel samples contained ∼40 μg of protein, and were visualized with Coomassie blue. Grl1p is marked by ∗; the prominent band migrating near 15 kDa in lanes 2–5 is chymotrypsin. Molecular weight standards are indicated.