Journal: Nature communications

Article Title: Transport and inhibition of the sphingosine-1-phosphate exporter SPNS2.

doi: 10.1038/s41467-025-55942-7

Figure Lengend Snippet: Fig. 2 | DDM binding pocket of SPNS2. a Bound n-dodecyl-β-D-maltopyranoside within the SPNS2 structure determined in DDM. Positive difference density of the weighted Fo −Fc difference map at 16σ and the DDM model are shown as blue surface and DDM cyan sticks, respectively. b Cross section of the SPNS2 structure, viewed from the plane of the membrane. c SPNS2 coordinates DDM through van der Waals contacts in the pocket and hydrogen bonds in the central cavity. SPNS2-

Article Snippet: Briefly, codon-optimized sequences of wild type and mutant SPNS2 in pDONR221 (Addgene #132307) were subcloned into a modified pJTI R4 DEST CMV TO pA plasmid (Thermo Fisher Scientific) containing Twin-Strep-Tag (IBA Lifesciences) and HA epitopes (SH tag) at the N or the C terminus of SPNS2 as indicated.

Techniques: Binding Assay, Membrane

Journal: Nature communications

Article Title: Transport and inhibition of the sphingosine-1-phosphate exporter SPNS2.

doi: 10.1038/s41467-025-55942-7

Figure Lengend Snippet: Fig. 3 | Substrate and inhibitor binding by SPNS2. a Sphingosine-1-phosphate head group interactions with SPNS2 in simulation cluster 1. Carbons of SPNS2 side chains and FTY720-P are shown in purple and cyan, respectively. Interaction types are annotated by PLIP77. b FTY720-P interactions with SPNS2 in simulation cluster 1. c Inhibition of S1P export by 33p measured by S1PR3-coupled export assay after

Article Snippet: Briefly, codon-optimized sequences of wild type and mutant SPNS2 in pDONR221 (Addgene #132307) were subcloned into a modified pJTI R4 DEST CMV TO pA plasmid (Thermo Fisher Scientific) containing Twin-Strep-Tag (IBA Lifesciences) and HA epitopes (SH tag) at the N or the C terminus of SPNS2 as indicated.

Techniques: Binding Assay, Inhibition

Journal: Nature communications

Article Title: Transport and inhibition of the sphingosine-1-phosphate exporter SPNS2.

doi: 10.1038/s41467-025-55942-7

Figure Lengend Snippet: Fig. 5 | Pathogenic mutations at essential locations in the SPNS2 structures. a Locations of pathogenic SPNS2 mutations within the structure. Loop 7-8 is not resolved in the structure, and the approximate location of Pro356 is indicated by a magenta sphere with a dotted edge. b The pathogenic mutation ΔS319 within the C-domain. c Asp163 in SPNS2 is within the conserved MFS motif A and forms hydrogen bonds specific to the outward-facing conformation (PDB: 8EX5). Hydrogen bonds are shown as dotted lines. d S1P export activity by pathogenic

Article Snippet: Briefly, codon-optimized sequences of wild type and mutant SPNS2 in pDONR221 (Addgene #132307) were subcloned into a modified pJTI R4 DEST CMV TO pA plasmid (Thermo Fisher Scientific) containing Twin-Strep-Tag (IBA Lifesciences) and HA epitopes (SH tag) at the N or the C terminus of SPNS2 as indicated.

Techniques: Mutagenesis, Activity Assay