Molecular phenotyping of the surfaceome of migratory chondroprogenitors and mesenchymal stem cells using biotinylation, glycocapture and quantitative LC-MS/MS proteomic analysis
Molecular phenotyping of the surfaceome of migratory chondroprogenitors and mesenchymal stem cells using biotinylation, glycocapture and quantitative LC-MS/MS proteomic analysis
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DOI：
10.1038/s41598-019-44957-y
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发表时间：
2019-06-21
影响因子：
4.6
通讯作者：
Mobasheri, Ali
中科院分区：
文献类型：
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作者：
Matta, Csaba;Boocock, David J.;Mobasheri, Ali
细胞表面蛋白的补体，统称为表面体，是正常分化过程和骨关节炎（OA）等病理发展的有用指标。我们使用生化和蛋白质组学工具来探索来自人类OA膝关节软骨的软骨源性祖细胞（CPC）的表面体并定义其生物标志物。这些细胞具有巨大的治疗潜力，但其未开发的生物学限制了它们的临床应用。我们采用生物素化结合糖捕获和高通量霰弹枪蛋白质组学来确定人骨髓间充质干细胞（MSCs）和人cpc的表面蛋白质组。我们从MSCs和CPCs中制备了细胞表面蛋白富集组分，然后使用蛋白质组学方法比较和评估未分化的MSCs和CPCs之间的蛋白质变化。共鉴定出1256种蛋白，其中791种（63%）为质膜、细胞表面或细胞外基质蛋白。对构成表面小体的蛋白质进行了注释和分类。我们的结果首次提供了与软骨生物学和OA相关的两种密切相关的细胞类型表面体的定量蛋白质组学数据库。这些结果可能为软骨形成分化过程中表面体的转化和骨性关节炎发展过程中的表型变化提供新的见解。
The complement of cell surface proteins, collectively referred to as the surfaceome, is a useful indicator of normal differentiation processes, and the development of pathologies such as osteoarthritis (OA). We employed biochemical and proteomic tools to explore the surfaceome and to define biomarkers in chondrogenic progenitor cells (CPC) derived from human OA knee articular cartilage. These cells have great therapeutic potential, but their unexplored biology limits their clinical application. We performed biotinylation combined with glycocapture and high throughput shotgun proteomics to define the surface proteome of human bone marrow mesenchymal stem cells (MSCs) and human CPCs. We prepared cell surface protein-enriched fractions from MSCs and CPCs, and then a proteomic approach was used to compare and evaluate protein changes between undifferentiated MSCs and CPCs. 1256 proteins were identified in the study, of which 791(63%) were plasma membrane, cell surface or extracellular matrix proteins. Proteins constituting the surfaceome were annotated and categorized. Our results provide, for the first time, a repository of quantitative proteomic data on the surfaceome of two closely related cell types relevant to cartilage biology and OA. These results may provide novel insights into the transformation of the surfaceome during chondrogenic differentiation and phenotypic changes during OA development.