ClC-4 is a member of the ClC family of chloride channels and is localized to the apical brush border membrane of intestinal enterocytes and to endosomes. Currently little is known about its regulation. A nucleotide requirement has been documented although the underlying mechanism has not been identified. We tested the hypothesis that ClC-4 could be phosphorylated by PKA or PKC. As well, we tested the hypothesis that ClC-4 directly binds nucleotides. Using purified peptides incorporating the intracellular N- and C-terminal regions, as well as lysates from ClC-4 transfected cell lines we showed that both peptides could be phosphorylated in a PKA-dependent manner, though the full-length protein could not. However, precipitation of ClC-4 with ATP Sepharose beads revealed that ClC-4 could bind to ATP in a magnesium-dependent manner and that this binding could be competitively inhibited. Future studies will determine the impact of this interaction on ClC-4 function in the plasma membrane and in endosomes.