ClC-5, an endosomal Cl(-)/H(+)
antiporter that is mutated in
Dent disease, is essential for endosomal acidification and re-uptake of small molecular weight
proteins in the renal proximal tubule. Eukaryotic
chloride channels (CLCs) contain two cytoplasmic CBS domains, motifs present in different
proteins, the function of which is still poorly understood. Structural studies have shown that ClC-5 can bind to
ATP at the interface between the CBS domains, but so far the potential functional consequences of
nucleotide binding to ClC-5 have not been investigated. Here, we show that the direct application of
ATP,
ADP and
AMP in inside-out patch experiments potentiates the current mediated by ClC-5 with similar affinities. The
nucleotides increase the probability of ClC-5 to be in an active, transporting state. The residues Tyr 617 and Asp 727, but not Ser 618, are crucial for the potentiation. These results provide a mechanistic and structural framework for the interpretation of
nucleotide regulation of a CLC transporter.