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. 2001 Apr 10;98(9):5055–5060. doi: 10.1073/pnas.081026598

Figure 2.

Figure 2

Activation of Easter by Snake when all three zymogens are coexpressed. Western blots with anti-Easter antibody to detect Easter in cell pellet (A and B) and culture medium (A and C) after transfection of S2 cells. (A) EAS-A is cleaved from zymogen form (z) to 35-kDa C-terminal product (c) when expressed with GD + SNK zymogens (lane 4 or 8), but not when expressed with only GD (lane 2 or 6) or SNK (lane 3 or 7). (B) When expressed with GD + SNK zymogens, EAS-A is cleaved to generate a 35-kDa product (c, lane 4) that migrates similarly to EAΔN, the functional Easter catalytic domain (lane 2); wild-type EA is cleaved to generate predominantly a smaller product (c′, lane 3). Double catalytic and zymogen-activation mutant EAR-V,S-A is not cleaved (lane 5). (C) EAS-A is cleaved when expressed with preactivated SNK (SNKΔN; lane 2) but not when expressed with preactivated GD (GDΔN1; lane 1).