血浆来源的细胞外囊泡的纯化和磷酸化蛋白质组学分析。
Purification and Phosphoproteomic Analysis of Plasma-Derived Extracellular Vesicles.
机构信息
Tymora Analytical Operations, West Lafayette, IN, USA.
出版信息
Methods Mol Biol. 2022;2504:147-156. doi: 10.1007/978-1-0716-2341-1_11.
Abstract
A successful phosphoproteomics analysis of extracellular vesicles (EVs) requires a unique approach, fine-tuned to address the challenges that have plagued plasma-based biomarker discovery. Here, I detail a procedure, which combines EVtrap-based high-recovery EV isolation, phase-transfer surfactant method for protein extraction, and PolyMAC-based enrichment of phosphopeptides. The combination of these methods provides a highly effective strategy for EV-based phosphoproteome analysis and leads to the discovery of novel phospho-markers previously undetectable.
摘要
要成功分析细胞外囊泡（EVs）的磷酸化蛋白质组学，需要一种独特的方法，该方法需要经过精心调整，以解决困扰基于血浆的生物标志物发现的挑战。在这里，我详细介绍了一种程序，该程序结合了 EVtrap 高回收率 EV 分离、相转移表面活性剂法蛋白提取和 PolyMAC 富集磷酸肽。这些方法的组合为基于 EV 的磷酸蛋白质组学分析提供了一种非常有效的策略，并发现了以前无法检测到的新型磷酸标记物。
相似文献
1
Purification and Phosphoproteomic Analysis of Plasma-Derived Extracellular Vesicles.血浆来源的细胞外囊泡的纯化和磷酸化蛋白质组学分析。
Methods Mol Biol. 2022;2504:147-156. doi: 10.1007/978-1-0716-2341-1_11.
2
Isolation and Identification of Plasma Extracellular Vesicles Protein Biomarkers.血浆细胞外囊泡蛋白生物标志物的分离与鉴定。
Methods Mol Biol. 2023;2660:207-217. doi: 10.1007/978-1-0716-3163-8_14.
3
Highly Efficient Phosphoproteome Capture and Analysis from Urinary Extracellular Vesicles.高效从尿液细胞外囊泡中捕获和分析磷酸化蛋白质组。
J Proteome Res. 2018 Sep 7;17(9):3308-3316. doi: 10.1021/acs.jproteome.8b00459. Epub 2018 Aug 17.
4
Plasma-Derived Extracellular Vesicle Phosphoproteomics through Chemical Affinity Purification.通过化学亲和纯化的血浆来源细胞外囊泡磷酸蛋白质组学
J Proteome Res. 2020 Jul 2;19(7):2563-2574. doi: 10.1021/acs.jproteome.0c00151. Epub 2020 May 28.
5
Profiling Phosphoproteome Landscape in Circulating Extracellular Vesicles from Microliters of Biofluids through Functionally Tunable Paramagnetic Separation.通过功能可调谐的顺磁分离从微升生物流体中循环细胞外囊泡中进行磷酸化蛋白质组特征分析。
Angew Chem Int Ed Engl. 2023 Jul 17;62(29):e202305668. doi: 10.1002/anie.202305668. Epub 2023 Jun 7.
6
Phosphoproteome analysis of cerebrospinal fluid extracellular vesicles in primary central nervous system lymphoma.原发性中枢神经系统淋巴瘤脑脊液细胞外囊泡的磷酸化蛋白质组分析。
Analyst. 2023 Jul 26;148(15):3594-3602. doi: 10.1039/d3an00670k.
7
Phosphoproteins in extracellular vesicles as candidate markers for breast cancer.细胞外囊泡中的磷蛋白作为乳腺癌的候选标志物。
Proc Natl Acad Sci U S A. 2017 Mar 21;114(12):3175-3180. doi: 10.1073/pnas.1618088114. Epub 2017 Mar 7.
8
Proteomics and Phosphoproteomics of Circulating Extracellular Vesicles Provide New Insights into Diabetes Pathobiology.循环细胞外囊泡的蛋白质组学和磷酸化蛋白质组学为糖尿病发病机制提供了新的见解。
Int J Mol Sci. 2022 May 21;23(10):5779. doi: 10.3390/ijms23105779.
9
The proteome and phosphoproteome of circulating extracellular vesicle-enriched preparations are associated with characteristic clinical features in type 1 diabetes.循环细胞外囊泡富集制剂的蛋白质组和磷酸化蛋白质组与 1 型糖尿病的特征性临床特征相关。
Front Endocrinol (Lausanne). 2023 Jul 28;14:1219293. doi: 10.3389/fendo.2023.1219293. eCollection 2023.
10
Sequential phosphoproteomics and N-glycoproteomics of plasma-derived extracellular vesicles.血浆衍生细胞外囊泡的顺序磷酸化蛋白质组学和 N-糖蛋白质组学。
Nat Protoc. 2020 Jan;15(1):161-180. doi: 10.1038/s41596-019-0260-5. Epub 2019 Dec 20.
引用本文的文献
1
Extracellular vesicle proteomics and phosphoproteomics identify pathways for increased risk in patients hospitalized with COVID-19 and type 2 diabetes mellitus.细胞外囊泡蛋白质组学和磷酸化蛋白质组学鉴定 COVID-19 住院患者和 2 型糖尿病患者风险增加的途径。
Diabetes Res Clin Pract. 2023 Mar;197:110565. doi: 10.1016/j.diabres.2023.110565. Epub 2023 Feb 1.
本文引用的文献
1
Plasma-Derived Extracellular Vesicle Phosphoproteomics through Chemical Affinity Purification.通过化学亲和纯化的血浆来源细胞外囊泡磷酸蛋白质组学
J Proteome Res. 2020 Jul 2;19(7):2563-2574. doi: 10.1021/acs.jproteome.0c00151. Epub 2020 May 28.
2
Liquid Biopsy as Novel Tool in Precision Medicine: Origins, Properties, Identification and Clinical Perspective of Cancer's Biomarkers.液体活检作为精准医学中的新型工具：癌症生物标志物的起源、特性、识别及临床前景
Diagnostics (Basel). 2020 Apr 13;10(4):215. doi: 10.3390/diagnostics10040215.
3
Sequential phosphoproteomics and N-glycoproteomics of plasma-derived extracellular vesicles.血浆衍生细胞外囊泡的顺序磷酸化蛋白质组学和 N-糖蛋白质组学。
Nat Protoc. 2020 Jan;15(1):161-180. doi: 10.1038/s41596-019-0260-5. Epub 2019 Dec 20.
4
The current status of the clinical utility of liquid biopsies in cancer.液体活检在癌症临床应用中的现状。
Expert Rev Mol Diagn. 2019 Nov;19(11):1031-1041. doi: 10.1080/14737159.2019.1664290. Epub 2019 Sep 8.
5
Applications of liquid biopsies for cancer.液体活检在癌症中的应用。
Sci Transl Med. 2019 Aug 28;11(507). doi: 10.1126/scitranslmed.aay1984.
6
Diagnostic and prognostic impact of cell-free DNA in human cancers: Systematic review.游离 DNA 在人类癌症中的诊断和预后影响：系统评价。
Mutat Res Rev Mutat Res. 2019 Jul-Sep;781:100-129. doi: 10.1016/j.mrrev.2019.05.002. Epub 2019 May 11.
7
Highly Efficient Phosphoproteome Capture and Analysis from Urinary Extracellular Vesicles.高效从尿液细胞外囊泡中捕获和分析磷酸化蛋白质组。
J Proteome Res. 2018 Sep 7;17(9):3308-3316. doi: 10.1021/acs.jproteome.8b00459. Epub 2018 Aug 17.
8
Phosphoproteins in extracellular vesicles as candidate markers for breast cancer.细胞外囊泡中的磷蛋白作为乳腺癌的候选标志物。
Proc Natl Acad Sci U S A. 2017 Mar 21;114(12):3175-3180. doi: 10.1073/pnas.1618088114. Epub 2017 Mar 7.
9
Characterization of Human Cancer Cell Lines by Reverse-phase Protein Arrays.利用反相蛋白质阵列对人癌细胞系进行表征
Cancer Cell. 2017 Feb 13;31(2):225-239. doi: 10.1016/j.ccell.2017.01.005.
10
Multiplatform-based molecular subtypes of non-small-cell lung cancer.基于多平台的非小细胞肺癌分子亚型
Oncogene. 2017 Mar;36(10):1384-1393. doi: 10.1038/onc.2016.303. Epub 2016 Oct 24.
Zotero 插件