Fig. 3: Engineering N-terminal CPD to facilitate the desired cleavage site. | Communications Biology

Fig. 3: Engineering N-terminal CPD to facilitate the desired cleavage site.

From: Facile expression of proteins with desired N-terminal amino acid via an engineered cysteine protease domain

Fig. 3: Engineering N-terminal CPD to facilitate the desired cleavage site.

a The design of the N-terminus CPD engineering. b SDS-PAGE analysis of the InsP6-induced cleavage efficacy of Fusion 2-6. Conditions: 200 μM of InsP6 in 50 mM Tris-HCl, pH 7.5, 37 °C, 3 h. SDS-PAGE (c) and LC-MS (d) analysis of the InsP6-induced cleavage of Fusion 2, 5, 7, and 8, 16 h. SDS-PAGE (e) and LC-MS (f) analysis of the InsP6 induced cleavage of Fusion 8, 9, and 10, 16 h. g SDS-PAGE analysis of Fusion 9 mutants CPD-A/GLH-NbHer2 or CPD-A/GLK-NbHer2, 8 h. Conditions for CPD cleavage: 1 mM of InsP6 in 50 mM Tris-HCl, pH 7.5, 37 °C. * indicates a half or third molecular weight of CPDs.

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