Analysis of the Oligomeric Structure of the Motor Protein Prestin

摘要: Prestin, a member of the solute carrier family 26, is expressed in basolateral membrane outer hair cells. This protein provides molecular basis for cell somatic electromotility, which crucial frequency selectivity and sensitivity mammalian hearing. It has long been known that there are abundantly approximately 11-nM particles present membrane. These were hypothesized to be motor proteins drive electromotility. Because calculated size prestin monomer too small form an particle, possibility oligomerization was examined. We investigated possible quaternary structures by lithium dodecyl sulfate-PAGE, perfluoro-octanoate-PAGE, membrane-based yeast two-hybrid system, chemical cross-linking experiments. obtained from different host or native cells, resistant dissociation sulfate behaves as stable oligomer on sulfate-PAGE. In homo-oligomeric interactions between prestin-bait/prestin-prey suggest molecules can associate with each other. Chemical experiments, perfluoro-octanoate-PAGE/Western blot, affinity purification experiments all indicate exists higher order oligomer, such tetramer, prestin-expressing yeast, lines Our data using hydrophobic hydrophilic reducing reagents dimer connected disulfide bond embedded core. may act building block producing oligomers cell's