Reproducible Determination of High-Density Lipoprotein Proteotypes
Reproducible Determination of High-Density Lipoprotein Proteotypes
复制标题
DOI：
10.1021/acs.jproteome.1c00429
复制
发表时间：
2021-10-22
影响因子：
4.4
通讯作者：
Wollscheid, Bernd
中科院分区：
文献类型：
--
作者：
Goetze, Sandra;Frey, Kathrin;Wollscheid, Bernd
高密度脂蛋白（HDL）是血液循环中含有许多不同蛋白质、脂质和RNA的多分子颗粒的异质混合物。基于质谱的蛋白质型分析的最新进展显示了在大型患者队列中分析蛋白质型的前景。为了创建实现这些数据独立采集（DIA）策略所需的光谱库，从具有多种生理HDL状态的300多名患者的血浆中分离HDL。建立了由296种蛋白质组和超过786种肽型组成的HDL蛋白质组谱库，并将DIA策略的性能作为检测健康个体与患有2型糖尿病和/或冠心病的患者队列之间HDL蛋白质型差异的基准。使用生成的光谱库对数据进行生物信息学询问表明，DIA方法能够进行稳健的HDL蛋白质型测定。通过使用光谱库实现的HDL肽型分析允许鉴定翻译后修饰，例如在APOA 1中，其可能影响HDL功能。从技术的角度来看，数据分析进一步表明，蛋白质和肽的量目前在不同的HDL蛋白质型之间比没有进一步富集的肽型更有区别。总之，基于DIA的HDL蛋白质分型使HDL蛋白质型的稳健数字化成为更大临床队列分析的基础。
High-density lipoprotein (HDL) is a heterogeneous mixture of blood-circulating multimolecular particles containing many different proteins, lipids, and RNAs. Recent advancements in mass spectrometrybased proteotype analysis show promise for the analysis of proteoforms across large patient cohorts. In order to create the required spectral libraries enabling these data-independent acquisition (DIA) strategies, HDL was isolated from the plasma of more than 300 patients with a multiplicity of physiological HDL states. HDL proteome spectral libraries consisting of 296 protein groups and more than 786 peptidoforms were established, and the performance of the DIA strategy was benchmarked for the detection of HDL proteotype differences between healthy individuals and a cohort of patients suffering from diabetes mellitus type 2 and/or coronary heart disease. Bioinformatic interrogation of the data using the generated spectral libraries showed that the DIA approach enabled robust HDL proteotype determination. HDL peptidoform analysis enabled by using spectral libraries allowed for the identification of post-translational modifications, such as in APOA1, which could affect HDL functionality. From a technical point of view, data analysis further shows that protein and peptide quantities are currently more discriminative between different HDL proteotypes than peptidoforms without further enrichment. Together, DIA-based HDL proteotyping enables the robust digitization of HDL proteotypes as a basis for the analysis of larger clinical cohorts.