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. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Biochimie. 2019 Jun 15;166:161–172. doi: 10.1016/j.biochi.2019.06.010

Figure 2. Purified proRgpB6His undergoes partial autoproteolytic processing with limited increase in enzymatic activity.

Figure 2.

ProRgpB6His (0.4 mg/ml) was incubated at room temperature in gingipain activity buffer with/without 5 mM DTT supplementation. At various time points, aliquots were removed and subjected to (A) SDS-PAGE electrophoresis, or (B) Western blot analysis with pAb anti-RgpB, mAb anti-6HisTag and pAb anti-CTD domain. (C) Active-site labelling using the biotinylated probe BiRK to detect active forms of partially processed proRgpB. Mature RgpB bearing a C-terminal HisTag (Veillard et al, 2015) was used as a control (lane M). (D) The activity of RgpB at each time point was determined with the chromogenic substrate L-BAPNA. Results are expressed as percentage of activity of the native RgpB incubated in the same condition (n = 3).