Immunolocalization of anion exchanger 2alpha in auditory sensory hair cells.
We have previously reported the isolation from a guinea pig organ of Corti cDNA library of a cDNA clone that encodes a novel isoform of the anion exchanger 2 (AE2) protein (Negrini, Rivolta, Kalinec and Kachar, 1995. Cloning of an organ of Corti anion exchanger 2 isoform with a truncated C-terminal domain. Biophys. Acta, 1236, 207-211). The deduced protein, named AE2alpha, has a conserved cytoplasmic domain and a short membrane domain with only two membrane spanning regions, as opposed to the fourteen present in the conventional AE2. Now, we are showing the immunolocalization and preliminary characterization of this protein using an antipeptide antibody specific for this novel AE2 isoform. In Western blots, this antibody binds to an approximately 89 kDa polypeptide that corresponds to a phosphorylated protein with serines as main phosphate acceptor residues. In immunofluorescence experiments, the antibody labels the stereocilia and the lateral wall of the outer hair cells and the stereocilia of the inner hair cells. Our results suggest that AE2alpha is a membrane-cytoskeletal linker in regions of the hair cell, where sensory transduction mechanisms take place.
Kalinec
F
F
Section on Structural Cell Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD 20850, USA.
Kalinec
G
G
Negrini
C
C
Kachar
B
B
D000595:Amino Acid Sequence; D000818:Animals; D027321:Anion Transport Proteins; D000918:Antibody Specificity; D017920:Antiporters; D015153:Blotting, Western; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D006199:Hair Cells, Auditory, Inner / Q000378:metabolism*; D018072:Hair Cells, Auditory, Outer / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D008858:Microscopy, Phase-Contrast; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000737:chemistry; D009475:Neurons, Afferent / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D064606:SLC4A Proteins; D015398:Signal Transduction / Q000235:genetics
D027321:Anion Transport Proteins; D017920:Antiporters; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D064606:SLC4A Proteins