Supplementary Figure 8: Cleavage comparison of T4L-PL5 protein.
From: SNAC-tag for sequence-specific chemical protein cleavage
Left, thrombin cleavage of T4L-LVPRGS-PL5. Cleavage conditions: protein 1 mg/mL, 50 mM Tris, pH 8.4, 200 mM NaCl, 0.5 mM CaCl2, 5 mM DPC, 22 °C, 16 h, thrombin: lane 2, 2 U/mL, lane 3, 6 U/mL, lane 4, 12 U/mL. One thrombin unit is defined as that amount of enzyme required to cleave 1 mg of a test protein when incubated in standard digest buffer at 20 °C for 16 h. Right, Ni2+ cleavage of T4L-GSHHW-PL5. Cleavage conditions: 1 mM NiCl2, 0.1 M CHES, 0.2 M NaCl, pH 8.2, 5 mM DPC, protein 1 mg/mL, 22 °C, 16 h. Full sequence of T4L-LVPRGS-PL5: MGSSHHHHHHSSGLHLRDAHMGNIFEMLRIDEGLRLKIYKDTEGYYTIGIGHLLTKSPSLNAAKSELDKAIGRNTNGVITKDEAEKLFNQDVDAAVRGILRNAKLKPVYDSLDAVRRAALINMVFQMGETGVAGFTNSLRMLQQKRWDEAAVNLAKSRWYNQTPNRAKRVITTFRTGTWDAYAAGGSGSLVPRGSDPEQLKWISFCXXXXXXXXXXXXFMLYRGRR (de novo designed membrane protein, full sequence to be released in a separate paper). Full sequence of T4L-GSHHW-PL5: MGSSHHHHHHSSGLHLRDAHMGNIFEMLRIDEGLRLKIYKDTEGYYTIGIGHLLTKSPSLNAAKSELDKAIGRNTNGVITKDEAEKLFNQDVDAAVRGILRNAKLKPVYDSLDAVRRAALINMVFQMGETGVAGFTNSLRMLQQKRWDEAAVNLAKSRWYNQTPNRAKRVITTFRTGTWDAYAAGGSGSGSHHWGSDPEQLKWISFCXXXXXXXXXXXXFMLYRGRR (de novo designed membrane protein, full sequence to be released in a separate paper).
