Extended Data Fig. 9: Qualitative gel-based analysis of purified proteins.
From: Molecular basis for the initiation of DNA primer synthesis
a. MpCAPP fragments and FL mutants. 1 µg of each purified MpCAPP variant was resolved on 12% SDS-PAGE and Coomassie stained. Note: FL WT, FL CC and ΔTPR fragment are fused to MBP. FL WT – full-length wild type WT, FL AxA – full-length D177A, D179A, ΔCTD – aa1-360, ΔTPR – aa100-546, PP – aa100-360, FL CC – full-length C462S, C464S. b. Mutants of MpCAPP PP domain. 1 µg of each purified MpCAPP PP mutant was resolved using 12% SDS-PAGE and Coomassie stained. AxA – D177A, D179A, RR – R142A, R143A, KK – K181A, K182A, KQN – K264A, Q265A, N274A. c. MsCAPP fragments. 2 µg of purified fragments of MsCAPP100-359 and MsCAPPaa111-328 were resolved using 12% SDS-PAGE and Coomassie stained. d. HsPrimPol full-length (HsFL) and HsPrimPol1-354 fragment (HsPP) and mutants. 2 µg of each purified variant was resolved using 12% SDS-PAGE and Coomassie stained. e. HsPP WT and mutants. 2 µg of each purified mutant was resolved on 12% SDS-PAGE and Coomassie stained. Note: AxA – D114A, E116A, RNR – R286A, N287A, R288A. f. Eukaryotic PrimPols. 2 µg of each purified PP was resolved using 15% SDS-PAGE and Coomassie stained. g. HsPri1. 1 µg of HsPri1 wild-type (HsPri1) or HsPri1 D109A, D111A, D306A (HsPri1AAA) was resolved using 12% SDS-PAGE and Coomassie stained.
