Abstract
In this study, mass spectrometry was used to explore the canine tear proteome. Tear samples were obtained from six healthy dogs, and one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (1D SDS-PAGE) was used as a first step to separate intact proteins into 17 bands. Each fraction was then trypsin digested and analysed by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS) to characterize the protein components in each fraction. In total, 125 tear proteins were identified, with MCA (Major Canine Allergen), Serum albumin, UPF0557 protein C10orf119 homolog, Collagen alpha-2(I) chain, Tyrosine -protein kinase Fer, Keratine type II cytoskeletal, Beta-crystallin B2, Interleukin-6 and Desmin occuring as the most confident ones with the highest scores. The results showed that the proteomic strategy used in this study was successful in the analysis of the dog tear proteome. To the best of our knowledge, this study is the first to report the comprehensive proteome profile of tears from healthy dogs by 1D SDS PAGE and MALDI-TOF. Data are available via ProteomeXchange with identifier PXD003124.
Highlights
Proteome is a set of proteins expressed in a given time by a given tissue
Few publications in the proteomics literature have evaluated the tear film of animals, especially dogs can be of particular interest, as they live in the same conditions and often suffer from diseases of similar aetiopathogenesis [15,16,17]
The aim of this study was to examine the proteome profile of dog tear samples through one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (1D SDS-PAGE) in combination with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS)
Summary
Proteome is a set of proteins expressed in a given time by a given tissue. Its name comes from a blend of proteins and genome. Proteomic analysis has become an important tool in biomedical and veterinary research [1,2,3]. A molecular analysis of tear film composition is a useful source of information for the diagnosis, prognosis and treatment of diseases of the eye, as well as systemic diseases in humans [8,9,10,11]. In addition to its clinical utility, the identification of biomarkers in tear film may be useful in developing new pharmacologically active molecules and diagnostic tests [12,13,14]. Despite well-developed veterinary ophthalmology research concerning dogs, reports on molecular studies of the tear film remains sparse, and in-
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