Physico-chemical characterization of the native and mutant protein cochlin, and its role in adult-onset hearing and balance loss

Abstract

This thesis investigates the role of the protein cochlin and its isoforms in DFNA9 autosomal dominant late onset senorineural loss and vestibular disorder by quantifying the concentration of cochlin in the inner fluid called perilymph. Through the use of affinity chromatography, high performance liquid chromatography, the Bradford assay, western blot analysis, and proteomics analysis by mass spectroscopy, I acquired data which suggests that the concentration of cochlin in the perilymph of bovine calf ears is at most on the order of 10- mg/ml. I also determined the total protein concentration of native bovine perilymph to be 2.1+0.2 mg/ml. Additionally, I discuss the theory of quasielastic light scattering along with its relevance to understanding the role of cochlin in DFNA9.