Winiarczyk Mateusz, Winiarczyk Dagmara, Banach Tomasz, Adaszek Lukasz, Madany Jacek, Mackiewicz Jerzy, Pietras-Ozga Dorota, Winiarczyk Stanislaw
Department of Vitreoretinal Surgery, Medical University of Lublin, 20-079 Lublin, Chmielna 1, Poland.
Department and Clinic of Animal Internal Diseases, University of Life Sciences, 20-612 Lublin, Głęboka 30, Poland.
PLoS One. 2015 Dec 23;10(12):e0144242. doi: 10.1371/journal.pone.0144242. eCollection 2015.
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 occurring 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.
在本研究中,采用质谱法探索犬泪液蛋白质组。从六只健康犬获取泪液样本,第一步使用一维十二烷基硫酸钠聚丙烯酰胺凝胶电泳(1D SDS-PAGE)将完整蛋白质分离成17条带。然后对每个组分进行胰蛋白酶消化,并通过基质辅助激光解吸/电离飞行时间串联质谱(MALDI-TOF-MS/MS)分析,以鉴定每个组分中的蛋白质成分。总共鉴定出125种泪液蛋白质,其中主要犬过敏原(MCA)、血清白蛋白、UPF0557蛋白C10orf119同源物、胶原蛋白α-2(I)链、酪氨酸蛋白激酶Fer、角蛋白II型细胞骨架、β-晶状体蛋白B2、白细胞介素-6和结蛋白得分最高,最为可靠。结果表明,本研究中使用的蛋白质组学策略成功地分析了犬泪液蛋白质组。据我们所知,本研究首次通过1D SDS PAGE和MALDI-TOF报告了健康犬泪液的综合蛋白质组图谱。数据可通过ProteomeXchange获得,标识符为PXD003124。