title: Proteomic analysis of the organ of corti using nanoscale liquid chromatography coupled with tandem mass spectrometry.
abstract: The organ of Corti (OC) in the cochlea plays an essential role in auditory signal transduction in the inner ear. For its minute size and trace amount of proteins, the identification of the molecules in pathophysiologic processes in the bone-encapsulated OC requires both delicate separation and a highly sensitive analytical tool. Previously, we reported the development of a high resolution metal-free nanoscale liquid chromatography system for highly sensitive phosphoproteomic analysis. Here this system was coupled with a LTQ-Orbitrap XL mass spectrometer to investigate the OC proteome from normal hearing FVB/N male mice. A total of 628 proteins were identified from six replicates of single LC-MS/MS analysis, with a false discovery rate of 1% using the decoy database approach by the OMSSA search engine. This is currently the largest proteome dataset for the OC. A total of 11 proteins, including cochlin, myosin VI, and myosin IX, were identified that when defective are associated with hearing impairment or loss. This study demonstrated the effectiveness of our nanoLC-MS/MS platform for sensitive identification of hearing loss-associated proteins from minute amount of tissue samples.
authors.affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA.|Department of Environmental, Agricultural & Occupational Health, University of Nebraska Medical Center, Omaha, NE 68198, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Peng
authors.affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA.
authors.forename: Miao
authors.initials: M
authors.lastname: Liu
authors.affiliation: Department of Biomedical Sciences, Creighton University, Omaha, NE 68178, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Pecka
authors.affiliation: Department of Biomedical Sciences, Creighton University, Omaha, NE 68178, USA.
authors.forename: Kirk W
authors.initials: KW
authors.lastname: Beisel
authors.affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA.|Mass Spectrometry and Proteomics Core Facility, University of Nebraska Medical Center, Omaha, NE 68198, USA.
authors.forename: Shi-Jian
authors.initials: SJ
authors.lastname: Ding
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D063990:Gene Ontology; D034381:Hearing Loss / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D009925:Organ of Corti / Q000378:metabolism*; D020543:Proteome / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D020543:Proteome