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.
Schmieder
S
S
Laboratoire de Physiologie des Membranes Cellulaires, CNRS UMR 6078, Université de Nice-Sophia Antipolis, Villefranche sur Mer, France.
Lindenthal
S
S
Ehrenfeld
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D018118:Chloride Channels / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes / Q000737:chemistry; D006031:Glycosylation; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014981:Xenopus
D000906:Antibodies; D018118:Chloride Channels; C115376:ClC-3 channel; D000939:Epitopes