Engineered full-length
PKCα and truncation constructs used
in this study. Shown are the domain layouts of the human PKCα
constructs engineered for this study, and WT PKCα for comparison
(native domains defined in Figure 1A). Each
engineered construct contained both a fluorescent labeling tag (the
11-residue ybbr target peptide to which the Sfp enzyme covalently
couples the fluor) and an affinity tag [the 9-residue HA peptide or
the maltose-binding protein (MBP)]. Truncation borders were as follows:
C1A–C1B–C2 (residues 26–294), C1B–C2 (residues
90–294), C2 (residues 157–294), C1A–C1B (residues
26–165), C1A (residues 26–100), and C1B (residues 90–165).