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Autophosphorylation of rNSP2. Purified rNSP2 was incubated with α-32P-labeled ATP or GTP or with γ-32P-labeled ATP or GTP for 1 h at 37°C. Afterwards, 10 U of CIP was added to some reaction mixtures, which were then incubated for 1 h at 37°C. Proteins in the samples were detected by SDS–12% PAGE and staining with Coomassie blue (A). 32P-labeled proteins in the gel shown in (A) were identified by autoradiography (B). CIP becomes radiolabeled due to its affinity for NTPs.
