Identification of a 120 kd hair-bundle myosin located near stereociliary tips.
By adapting to sustained stimuli, hair cells of the internal ear maintain their optimal sensitivity to minute displacements. Biophysical experiments have suggested that adaptation is mediated by a molecular motor, most likely a member of the myosin family. To provide direct evidence for the presence of myosin isozymes in hair bundles, we used photoaffinity labeling with vanadate-trapped uridine and adenine nucleotides to identify proteins of 120, 160, and 230 kd in a preparation of hair bundles purified from the bullfrog's sacculus. The photoaffinity labeling properties of these proteins, particularly the 120 kd protein, resembled those of other well-characterized myosins. A 120 kd hair-bundle protein was also recognized by a monoclonal antibody directed against a vertebrate myosin I isozyme. Immunofluorescence microscopy localized this protein near the beveled top edge of the hair bundle, the site of mechanoelectrical transduction and adaptation.
Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas 75235-9039.
P G
PG
Gillespie
M C
MC
Wagner
A J
AJ
Hudspeth
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D001345:Autoradiography; D002118:Calcium / Q000494:pharmacology; D002923:Cilia / Q000737:chemistry* / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006198:Hair Cells, Auditory / Q000737:chemistry* / Q000201:enzymology; D007527:Isoenzymes / Q000032:analysis / Q000302:isolation & purification; D008970:Molecular Weight; D009218:Myosins / Q000032:analysis / Q000302:isolation & purification*; D010761:Phosphorus Radioisotopes; D011892:Rana catesbeiana; D014544:Uridine Triphosphate / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D010761:Phosphorus Radioisotopes; D014638:Vanadates; D000255:Adenosine Triphosphate; D009218:Myosins; D002118:Calcium; D014544:Uridine Triphosphate