title: Cross-sample validation provides enhanced proteome coverage in rat vocal fold mucosa.
abstract: The vocal fold mucosa is a biomechanically unique tissue comprised of a densely cellular epithelium, superficial to an extracellular matrix (ECM)-rich lamina propria. Such ECM-rich tissues are challenging to analyze using proteomic assays, primarily due to extensive crosslinking and glycosylation of the majority of high M(r) ECM proteins. In this study, we implemented an LC-MS/MS-based strategy to characterize the rat vocal fold mucosa proteome. Our sample preparation protocol successfully solubilized both proteins and certain high M(r) glycoconjugates and resulted in the identification of hundreds of mucosal proteins. A straightforward approach to the treatment of protein identifications attributed to single peptide hits allowed the retention of potentially important low abundance identifications (validated by a cross-sample match and de novo interpretation of relevant spectra) while still eliminating potentially spurious identifications (global single peptide hits with no cross-sample match). The resulting vocal fold mucosa proteome was characterized by a wide range of cellular and extracellular proteins spanning 12 functional categories.
authors.affiliation: Department of Surgery, Division of Otolaryngology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, United States of America. welham@sugery.wisc.edu
authors.forename: Nathan V
authors.initials: NV
authors.lastname: Welham
authors.forename: Masaru
authors.initials: M
authors.lastname: Yamashita
authors.forename: Seong Hee
authors.initials: SH
authors.lastname: Choi
authors.forename: Changying
authors.initials: C
authors.lastname: Ling
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D007820:Laryngeal Mucosa / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D010455:Peptides / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D014827:Vocal Cords / Q000378:metabolism*
chemical_list: D016326:Extracellular Matrix Proteins; D010455:Peptides; D020543:Proteome