1. RNaseB was partially digested with subtilisin, yielding a preparation that contained undigested RNaseB and the faster migrating RNaseBS‐Prot (lane 1).
RNaseB was partially digested with subtilisin, yielding a preparation that contained undigested RNaseB and the faster migrating RNaseBS‐Prot (lane 1). Digestion with Endo H deglycosylated both forms of the RNaseB (lane 2), whereas Png1 deglycosylated only the RNaseBS‐Prot (lane 3). RNaseB reconstituted with S‐peptide is susceptible to deglycosylation by Endo H (lane 4), but resistant to deglycosylation by Png1 (lane 5). The activity of Endo H is not reduced in the absence of DTT (lane 6), whereas Png1 activity shows a marked reduction in activity (lane 7). Endo H: 25 U; Png1: 42 U. Total RNaseB concentration was 5 μg in a final volume of 30 μl. Reactions were incubated at 20–23°C for 16 h.
Christian Hirsch et al. EMBO Rep. 2004;5:
© as stated in the article, figure or figure legend