Fig. 1: Biochemical functionality of agonistic, hormetic and antagonistic effects of HM model compounds in vitro.
From: Enabling systemic identification and functionality profiling for Cdc42 homeostatic modulators
Cdc42 HM model compounds regulate Cdc42 GTP loading and its GEF-mediated activation. ZCL278 and ZCL279 (at low concentrations) increase Cdc42 GTP loading while ZCL367 decreases Cdc42 GTP loading, A ZCL278, C ZCL279 and E ZCL367 without GEF. ZCL278, ZCL279 (at high concentrations) and ZCL367 decrease GEF-mediated Cdc42 GTP loading, B ZCL278, D ZCL279 and F ZCL367 with GEF. HM model compounds act through GEF like mechanism, The amount of GTP loading on Cdc42 for ITSN as a full agonist were comparable to those for HMs in different concentrations (0.01–1 µM) of G ZCL278, H ZCL279 and I ZCL367 respectively. Control for all samples without ITSN was DMSO and Cdc42, for all samples with GEF was DMSO, Cdc42 and ITSN. All data are presented as mean ± SEM from duplicates or triplicates in at least two independent experiments. ANOVA compared treatments to their respective control (*p < 0.04, **p < 0.005, ***p < 0.0007, ****p < 0.0001). The GEF reaction was performed using the relative fluorescence unit (RFU) intensity of Mant-GTP exchange that monitors GDP release during the exchange reaction after treatment by different concentrations (0.1–100 µM) of ZCL278, ZCL279 and ZCL367 on Cdc42 in absence or presence of ITSN.
