Figure 8.

Zymogram analysis of glycogen/starch synthase activities. Total proteins of crude extracts of the wild-type strain, the 187G11 mutant strain, and the recombinant proteins GlgA1, GlgA2, and GlgA2* were separated by native PAGE containing glycogen (A) or without glycogen and then electrotransferred onto native PAGE gels containing 0.6% (p/v) glycogen (B). The native gels were then incubated with or without 3 mm ADP-Glc. Glycogen/starch synthase activities are seen after iodine staining as dark activity bands. The wild-type glycogen/starch synthase and the recombinant protein GlgA2 comigrate when glycogen is absent from the zymogram gels (B), while in the presence of glycogen on the zymogram gels, the wild-type enzyme migrates slower then the recombinant protein GlgA2 (A). Numbered bars are as follows: 1, 187G11; 2, GlgA2*; 3, the wild type; 4, GlgA2; and 5, GlgA1.