SDS/PAGE of A. tumefaciens GS-reaction products. Purified GS was incubated with ADP-[14C]Glc under unprimed initiation reaction conditions, and 10% TCA-insoluble material was submitted to 10% SDS/PAGE. (A) Coomassie brilliant blue staining. (B) Autoradiography of the gel shown in A.(C) Reactions were carried out as described for A and stopped by heating tubes at 65°C for 15 min, the contents were treated with α-amylase, and precipitates insoluble in 10% TCA were submitted to 10% SDS/PAGE. Lane 1, enzymatically treated; lane 2, untreated control. The arrow indicates GS migrating position. (D and E) Reactions were carried out and stopped as described for C and submitted to proteinase K treatment, and precipitates insoluble in 10% TCA were run on 10% SDS/PAGE. (D) Coomassie brilliant blue staining. (E) Autoradiography of the gel shown in D. Lanes 1, enzymatically treated samples; lanes 2, untreated controls. For further details see Materials and Methods.