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Image Search Results
Journal: Nature
Article Title: Capturing a rhodopsin receptor signalling cascade across a native membrane
doi: 10.1038/s41586-022-04547-x
Figure Lengend Snippet: a , Following absorption of a photon of light hv (1), 11 -cis- retinal of rho isomerizes to the all- trans isomer (2). The activated states of rho (rho*) engage with transducin (G t ), consisting of Gα t •GDPβγ t , and exchange GDP for GTP. G t dissociates to form Gα t •GTP and Gβγ t ; loss of retinal from rho* leads to the formation of opsin (3). α-Subunits of G t interact with the γ-subunits in the PDE6 enzyme, with γ-subunits undergoing a conformational change, relieving inhibition and thereby activating PDE6 to cause hydrolysis of cGMP (4). Depletion of cGMP then closes the ion channel and the ‘dark current’ is terminated (5). The resulting change in the membrane potential produces the sensation of light. b , Disc membranes of rod cells are homogenized to form a heterogeneous distribution of vesicles that are introduced directly into the mass spectrometer (MS). The spectrum shown was recorded using the parameters stated above, which led to the dissociation of lipids from proteins. c , An LED light source is configured to apply timed light intervals before the electrospray ionization of the vesicles into an Orbitrap mass spectrometer under the conditions noted. HCD, higher-energy collisional dissociation. d , Following the addition of a soluble fraction containing PDE6 and G t , all proteins along the signalling pathway were detected. The proteins were ejected intact as rho/opsin (red), trimeric G t •GDP (cyan) and tetrameric PDE6•cGMP (pink). The mass spectrum shown from m / z 4,000 to m / z 8,000 represents the raw data. The experiment was repeated three times.
Article Snippet: The evaporated lipid mixture was dissolved in a 50 µl buffer and sonicated for 10 min. For liquid chromatography–MS/MS analysis, lipids were loaded onto a C8 column (Acclaim PepMap 100, C8, inner diameter of 75 µm, particle size of 3 µm, length of 150 mm; Thermo Scientific) using a Dionex UltiMate 3000 RSLC nano System connected to an
Techniques: Inhibition, Mass Spectrometry