Design of Retrocyclin Constructs
(A) Shows a schematic of the four constructs (R1, R3, A1, and A3) used for stable transfections along with native retrocyclin cDNA. All constructs have two termination codons at the end to ensure read-fidelity (red). Constructs A1 and A3 contain additional downstream residues (orange), whereas constructs R1 and R3 lack them. The two arrows indicate the position at which the two site-directed mutagenesis (⊗17Q and R70K) were performed.
(B) Shows the three possible mature retrocyclin peptides that could be formed from the constructs, homodimers of R1 or A1 encoding RC-100 (wild-type retrocyclin), heterodimers of A1 and A3 or R1 and R3 encoding RC-101 (single lysine congener), and homodimers of R3 or A3 encoding RC-101_2K (double lysine congener).