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Custom Vector Construction> | … Codon-optimized HnSpns (GenScript) was cloned into pET19b vector encoding an N-terminal 10-… Codon-optimized MsSpns (GenScript) was cloned into pET19b vector encoding an N-… | Get A Quote |
Spinster (Spns) lipid transporters are critical for transporting sphingosine-1-phosphate (S1P) across cellular membranes. In humans, Spns2 functions as the main S1P transporter in endothelial cells, making it a potential drug target for modulating S1P signaling. Here, we employed an integrated approach in lipid membranes to identify unknown conformational states of a bacterial Spns from Hyphomonas neptunium (HnSpns) and to define its proton- and substrate-coupled conformational dynamics. Our systematic study reveals conserved residues critical for protonation steps and their regulation, and how sequential protonation of these proton switches coordinates the conformational transitions in the context of a noncano... More