Laboratory of Proteomics and Mass Spectrometry and Department of Experimental and Clinical Medicine, "Magna Graecia" University of Catanzaro, Catanzaro, Italy.
Electrophoresis. 2011 Mar;32(6-7):686-95. doi: 10.1002/elps.201000592. Epub 2011 Feb 21.
The proteome of quiescent human platelets was analyzed by a shotgun proteomics approach consisting of enzymatic digestion, peptide separation based on isoelectric point by the use of OFFgel fractionation and, finally, RP nanoscale chromatography coupled to MS/MS detection (nano-LC-MS/MS). OFFgel fractionation in the first dimension was effective in providing an additional dimension of separation, orthogonal to RP nano-LC, thus generating an off-line multidimensional separation platform that proved to be robust and easy to set up. The analysis identified 1373 proteins with high confidence (false discovery rate<0.25%). The core set of 1373 human platelet proteins was investigated by Ingenuity Pathway Analysis software from which ten canonical pathways and eight networks have been validated, to suggest that platelets behave either as inflammatory or immune cells, and plasma membrane and cytoskeleton proteins play a fundamental role in their function. Moreover, toxicity pathway in agreement with network analysis, supports the concept that platelet life span is governed by an apoptotic mechanism.
通过一种包括酶解、基于等电点使用 OFFgel 分段的肽分离以及最后与 MS/MS 检测（纳升液相色谱-MS/MS）耦联的 shotgun 蛋白质组学方法分析了静止人血小板的蛋白质组。在第一维的 OFFgel 分段在提供与 RP 纳升 LC 正交的额外分离维度方面是有效的，从而生成了被证明是稳健且易于建立的离线多维分离平台。分析鉴定了 1373 种具有高可信度（错误发现率<0.25%）的蛋白质。通过 Ingenuity Pathway Analysis 软件研究了核心的 1373 个人血小板蛋白质组，从中验证了十个经典途径和八个网络，表明血小板表现为炎症或免疫细胞，并且质膜和细胞骨架蛋白在其功能中起着基本作用。此外，与网络分析一致的毒性途径支持血小板寿命受凋亡机制控制的概念。
