Active Prestin (PRES)

Prestin (PRES), encoded by the SLC26A5 gene, is a key motor protein exclusively expressed in outer hair cells (OHCs) of the mammalian cochlea. It mediates the unique "electromotility" of OHCs—rapid shape changes in response to electrical signals—which amplifies sound vibrations and enhances hearing sensitivity. As a member of the SLC26 anion transporter family, prestin uses voltage-dependent conformational shifts to drive OHC length changes, a process critical for fine-tuning auditory responses. Currently, there is no direct evidence of physical binding between prestin (PRES) and CFTR; their functional interactions in ion homeostasis remain to be clarified.Briefly, CFTR was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PRES-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CFTR pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately.Measured by its binding ability in a functional ELISA. When Recombinant PRES is lmmobilized at 2 µg/mL(100 uLwell), the concentration of CFTR that produces 50% optimal bindingresponse is found to be approximately 0.39287 µg/mL.