Fig. 6: MD simulations and stopped-flow measurements of GlpF and its mutant.
From: Narrowed pore conformations of aquaglyceroporins AQP3 and GlpF
a–d Initial and monitored distance between Val173(Cα) and Ala79(Cα) (a, b) and between Gly170(Cα) and Ala79(Cα) (c, d) in MD simulations for IGlpF/POPC. In the graphs of the monitored distance, the plots of the four chains of tetramers are shown (b, d). The loop D is represented by magenta (a, c). e Representative water distribution and the moved loop D in a snapshot for IGlpF/POPC during the MD simulation. The loop D and water molecules are represented by magenta and spheres, respectively. Initial (f) and monitored distance (g) between Val173(Cα) and Ala79(Cα) in MD simulations for I GlpF-ΔP177/POPC. In the graphs of the monitored distance, the plots of the four chains of tetramers are shown (g). The loop D is represented by magenta. h Water distribution in pores during MD simulations for two systems. The pore axes are aligned with z-axes of the cryo-EM structures. The origin of the pore axis is set at the z-coordinate of Cα atom of Asn68. i The measured protein/phospholipid weight ratio of GlpF and GlpF ΔP177 proteoliposomes. Averaged values ± SEM and individual data points are plotted (n = 4 independently reconstituted liposome samples from the same purified proteins). Data are statistically analysed using Student’s two-tailed unpaired t test. P value is indicated. j Stopped-flow measurements under NaCl hyperosmotic conditions for GlpF and GlpF ΔP177 proteoliposomes and empty liposomes. Averaged water permeability Pf values ± SEM and individual data points are plotted (n = 4 independently reconstituted liposome samples from the same purified proteins). Data are statistically analysed using multiple comparison Tukey test. P values are indicated. ***P < 0.001. k Stopped-flow measurements under glycerol gradient conditions for GlpF and GlpF ΔP177 proteoliposomes and empty liposomes. Averaged glycerol permeability Pgly values ± SEM and individual data points are plotted (n = 4 independently reconstituted liposome samples from the same purified proteins). Data are statistically analysed using multiple comparison Tukey test. P values are indicated. ***P < 0.001. Source data are provided as a Source Data file.
