In situ binding assay to detect Myosin-1c interactions with hair-cell proteins.
Myosin-1c is an unconventional myosin involved in hair-cell mechanotransduction, a process that underlies our senses of hearing and balance. To study the interaction of myosin-1c with other components of the hair-cell transduction complex, we have developed an in situ binding assay that permits visualization of myosin-1c binding to hair-cell proteins. In this chapter we describe in detail the methods needed for the expression and purification of recombinant myosin-1c fragments and their use in the in situ binding assay.
Phillips
Kelli R
KR
Sensory Neuroscience Research Center, Department of Biochemistry and Molecular Pharmacology, West Virginia University School of Medicine, Morgantown, WV, USA.
Cyr
Janet L
JL
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D001703:Biophysics / Q000379:methods; D002460:Cell Line; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D004594:Electrophysiology / Q000379:methods; D006198:Hair Cells, Auditory / Q000378:metabolism*; D006801:Humans; D007313:Insecta; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D024461:Myosin Type I / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D024461:Myosin Type I; C460367:MYO1C protein, human