Fig. 3: LYCHOS adopts an expanded conformation without sufficient cholesterol.
From: Structural insights into cholesterol sensing by the LYCHOS-mTORC1 pathway
a Cartoon representation of LYCHOSWT_C (cyan) and LYCHOSWT_E (yellow), superimposed based on the PLD domain of chain A in LYCHOSWT_C and chain B in LYCHOSWT_E. b Conformational displacements of the GLD relative to the PLD between LYCHOSWT_C (cyan) and LYCHOSWT_E (yellow). Cα atoms of residues used to measure displacements are shown in sticks and labeled. c Schematic representation of the displacement of the four residues highlighted in b. Dashed lines indicate distances between the Cα atoms within each state. Solid arrows indicate the absolute distance between Cα atoms of each residue between LYCHOSWT_E and LYCHOSWT_C. d, e The distance between Tyr551 and bilayer phospholipids is larger in LYCHOSWT_C when compared with LYCHOSWT_E. The bilayer phospholipids are shown in stick. Tyr551 is shown in stick and colored in magenta. The states were revealed by 1 μs molecular dynamics simulation. f Schematic of the FlAsH-BRET assay. Nluc inserted at intracellular loops (ICL2 or ICL5) of LYCHOS (transporter-like domain) and the FlAsH motif (CCPGCC) incorporated in ICL7, ICL8 or C-terminus of LYCHOS (GPCR-like domain). LYCHOSWT_E (gray) and LYCHOSWT_C (green) are shown from cytosolic view. g Detailed description of the Nluc and FlAsH motif incorporation sites at the ICLs of LYCHOS. h, i The maximal response of six intramolecular fluorescent arsenical hairpin (FlAsH)–bioluminescence resonance energy transfer biosensors of LYCHOS in response to cholesterol stimulation; The values are reported as the mean ± SEM of three biologically independent experiments (n = 3). j, k Concentration-dependent curves of LYCHOS FlAsH-BRET sensors in response to cholesterol (n  =  3 biologically independent experiments). l Cartoon representation of LYCHOSWT_C (cyan) and LYCHOSY57A_C (slate), superimposed based on the PLD domain. m Conformational displacements of the LYCHOSWT_C (cyan) bound CHS molecules (cyan) and LYCHOSY57A_C (slate) bound CHS molecules (green) relative to the PLD. Solid arrows indicate the absolute distance between carboxylate oxygen atoms of CHS molecules between LYCHOSWT_C and LYCHOSY57A_C. n Superimposition of the LYCHOSY57A_C structure with the map of LYCHOSY57A/R61A_E. The solid arrow indicates the translation of GLD.
