Extended Data Fig. 4: Purification and enzymatic activities of CorePro variants.
From: Substrate recognition and cleavage mechanism of the monkeypox virus core protease
a, TATV CorePro and c, MPXV CorePro I215K eluted as a single peak on size-exclusion chromatography. The purity as judged by SDS-PAGE was >90%. TATV CorePro and MPXV CorePro I215K were in the right lane of the corresponding SDS-PAGEs; the left lanes were WT MPXV CorePro. Both b, TATV CorePro and d, MPXV CorePro I215K cleaved S-G32, a fluorescent substrate derived from the second cleavage site of MPXV protein P25K. e, MPXV CorePro D258A and g, D248A eluted on a Superdex 200 increase size-exclusion column. The purity as judged by SDS-PAGE was >90%. Neither f, MPXV CorePro D258A nor h, D248A was able to cleave S-G32. i, A MPXV CorePro variant, which consists of residues 1–405, eluted as a single peak on Superdex 75 Increase size-exclusion column and demonstrated >90% purity on SDS-PAGE, and j, did not cleave S-G32. AU is the abbreviation for arbitrary unit. The enzymatic activity data in b and d were derived from n = 4 biological replicates and are presented as mean values ± s.d.
