Fig. 3: PGE2 recognition by SLCO2A1.
From: Structure and transport mechanism of the human prostaglandin transporter SLCO2A1
a, b The EM map of SLCO2A1 in complex with PGE2 (SLCO2A1-PGE2) embedded in LMNG micelle (gray) (a) and the cartoon representation of SLCO2A1-PGE2 (b). The PGE2 is colored in cyan. The N and C domain of SLCO2A1-PGE2 are colored in green and pink, respectively. c A cut-open electrostatic potential surface of PGE2 binding pocket. PGE2 is depicted in cyan sticks. The electrostatic potential surface was calculated in PyMOL (red to blue, −50 kT/e− to +50 kT/e−). d The electron microscopy densities of PGE2 and nearby residues. The EM densities for PGE2 and nearby residues were contoured at 5.8 σ. The density of PGE2 and nearby residues are shown in blue and orange meshes, respectively. e, f Detailed interactions between PGE2 and SLCO2A1. Residues involved in PGE2-binding are shown as sticks. Hydrogen bond interactions are indicated in orange dashed lines. Distances are indicated by black dashed lines. g 6-CF transport by HEK293F cells expressing SLCO2A1 WT or SLCO2A1 mutants involved in PGE2 binding. Cells expressing mCherry served as a control. Data were normalized to SLCO2A1 and are presented as mean ± s.e.m.; n = 3 independent replicates for mutants. n = 9 independent replicates for mCherry and SLCO2A1 WT. One-way ANOVA was used; ****P < 0.0001. h Concentration-dependent inhibition of 6-CF transport by PGE2 in HEK293F cells expressing SLCO2A1 mutants, S346A, S346F and W565F. Cells expressing mCherry served as a control. Data were normalized to net 6-CF transport activity of S346A, S346F or W565F without PGE2 and are presented as mean ± s.e.m.; For S346A, n = 6 independent replicates for log [PGE2 (μM)] = −3, n = 7 independent replicates for log [PGE2 (μM)] = −2, n = 3 independent replicates for all other conditions; For S346F, n = 3 independent replicates for log [PGE2 (μM)] = −1 or 0, n = 4 for all other conditions; For W565F, n = 7 independent replicates for log [PGE2 (μM)] = −3, n = 6 independent replicates for log [PGE2 (μM)] = −1, n = 3 independent replicates for all other conditions in W565F. Source Data are provided as a Source Data file.
