Trichler Shauna A, Bulla Sandra C, Mahajan Nandita, Lunsford Kari V, Pendarvis Ken, Nanduri Bindu, McCarthy Fiona M, Bulla Camilo
Department of Pathobiology and Population Medicine.
Department of Clinical Sciences and Animal Health Center, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS.
Vet Med (Auckl). 2014 Jun 21;5:1-9. doi: 10.2147/VMRR.S47127. eCollection 2014.
During platelet development, proteins necessary for the many functional roles of the platelet are stored within cytoplasmic granules. Platelets have also been shown to take up and store many plasma proteins into granules. This makes the platelet a potential novel source of biomarkers for many disease states. Approaches to sample preparation for proteomic studies for biomarkers search vary. Compared with traditional two-dimensional polyacrylamide gel electrophoresis systems, nonelectrophoretic proteomics methods that employ offline protein fractionation methods such as the differential detergent fractionation method have clear advantages. Here we report a proteomic survey of the canine platelet proteome using differential detergent fractionation coupled with mass spectrometry and functional modeling of the canine platelet proteins identified. A total of 5,974 unique proteins were identified from platelets, of which only 298 (5%) had previous experimental evidence of in vivo expression. The use of offline prefractionation of canine proteins by differential detergent fractionation resulted in greater proteome coverage as compared with previous reports. This initial study contributes to a broader understanding of canine platelet biology and aids functional research, identification of potential treatment targets and biomarkers, and sets a new standard for the resting platelet proteome.
在血小板发育过程中，血小板发挥多种功能所需的蛋白质存储在细胞质颗粒内。研究还表明，血小板能够摄取并将许多血浆蛋白存储到颗粒中。这使得血小板成为许多疾病状态下潜在的新型生物标志物来源。用于寻找生物标志物的蛋白质组学研究的样品制备方法各不相同。与传统的二维聚丙烯酰胺凝胶电泳系统相比，采用诸如差异去污剂分级分离法等离线蛋白质分级分离方法的非电泳蛋白质组学方法具有明显优势。在此，我们报告了一项利用差异去污剂分级分离结合质谱法对犬血小板蛋白质组进行的蛋白质组学调查，并对鉴定出的犬血小板蛋白质进行了功能建模。从血小板中总共鉴定出5974种独特蛋白质，其中只有298种（5%）有体内表达的先前实验证据。与先前的报告相比，通过差异去污剂分级分离对犬蛋白质进行离线预分级能够实现更大的蛋白质组覆盖范围。这项初步研究有助于更广泛地了解犬血小板生物学，并有助于功能研究、潜在治疗靶点和生物标志物的鉴定，为静息血小板蛋白质组设定了新的标准。
