Extended Data Fig. 5: cpLOV2 affords new surface areas to cage protein activity.
From: Circularly permuted LOV2 as a modular photoswitch for optogenetic engineering
Engineered STIM1-mediated activation of the ORAI Ca2+ channel was utilized to test the caging capability. Light-induced Ca2+ influx in HeLa cells was monitored by GCaMP6m. To create light-operated Ca2+ channel actuators (LOCCa), two effector domains (SOAR-I, STIM1336–486 or SOAR-II, STIM1344–442) were incorporated into LOV2 or cpLOV2 as indicated, in the presence or absence of Zdk2. Samples sizes: n = 25 and 30 cells for V1(left and right bars) from three independent biological replicates; V2, 49 and 50; V3, 25 and 25; V4, 42 and 25; V5, 25 and 25; V6, 27 and 61; V7, 31 and 25; V8, 46 and 46. Data are shown as mean ± s.e.m. Fmax, maximal fluorescent signals with photostimulation; F0, basal fluorescent signals in the dark.
