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. 2018 Mar 7;13(3):e0193947. doi: 10.1371/journal.pone.0193947

Fig 2. Single-Cys GCAP1 mutants retain the ability to regulate RetGC.

Fig 2

Membranes from HEK 293 cells expressing guanylyl cyclase RetGC1 were reconstituted with the purified recombinant myristoylated GCAP1 and assayed as described under Materials and Methods. (A) RetGC1 activity after reconstitution with 10 μM wild type, Cys-less GCAP1CL and single-Cys mutants measured in the presence of either EGTA (<10 nM free Ca2+) or 50 μM free Ca2+; mean average ± SD from 3 measurements. (B) Dose-dependence of the cyclase activation in the presence of EGTA by wild type GCAP1 (○), a Cys-less mutant GCAP1CL(●) or mutants containing a single Cys, GCAP1CL(T29C) (▲), GCAP1CL(E57C) (◇), GCAP1CL(E133C(◆), and GCAP1CL(E154C) (△); mean average ± SD from 3 independent experiments.