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DNA Sequencing> | The genes encoding VcINDY and MT5 were synthesized (GenScript, NJ) and cloned into a modified pET28b vector with an N-terminal cleavable deca-histidine tag. Mutations were introduced into the gene-encoding VcINDY by the QuikChange method (Agilent Technologies) and were confirmed by DNA sequencing. | Get A Quote |
Integral membrane proteins of the divalent anion/Na symporter (DASS) family translocate dicarboxylate, tricarboxylate or sulphate across cell membranes, typically by utilizing the preexisting Na gradient. The molecular determinants for substrate recognition by DASS remain obscure, largely owing to the absence of any substrate-bound DASS structure. Here we present 2.8-Å resolution X-ray structures of VcINDY, a DASS from Vibrio cholerae that catalyses the co-transport of Na and succinate. These structures portray the Na-bound VcINDY in complexes with succinate and citrate, elucidating the binding sites for substrate and two Na ions. Furthermore, we report the structures of a humanized variant of VcIN... More