Tissue-specific N-glycosylation of the ClC-3 chloride channel

A commercially available polyclonal antibody against a rClC-3/GST fusion protein was used in order to investigate the tissue distribution of the ClC-3 chloride channel protein. The antibody appeared to be specific to rClC-3 since no cross-reaction could be observed with rClC-4 or rClC-5 proteins when overexpressed in Xenopus oocytes. In mouse, mClC-3 was preferentially expressed in the central nervous system, intestine, and kidney. To a lower extent, mClC-3 protein was also detected in liver, lung, skeletal muscle, and heart. Surprisingly, the electrophoretic mobility of mClC-3 differed in the various tissues. After enzymatic digestion of N-linked oligosaccharide residues of membrane proteins from brain, intestine, and kidney, mClC-3 was found to migrate at its calculated molecular mass. This study provides important information regarding the specificity of the used antibody, indicates that ClC-3 is widely expressed in mouse, and that mClC-3 undergoes differential tissue-specific N-glycosylation.