PDF(Pubmed)
Abstract：
Over the past decades, proteomics has become increasingly important and a heavily discussed topic. The identification of intact proteins remains a major focus in this field. While most intact proteins are analyzed using high-resolution mass spectrometry, identifying them through low-resolution mass spectrometry continues to pose challenges. In our study, we investigated the capability of identifying various intact proteins using collision-induced dissociation (CID) and electron transfer without dissociation (ETnoD). Using myoglobin as our test protein, stable product ions were generated with CID, and the identities of the product ions were identified with ETnoD. ETnoD uses a short activation time (AcT, 5 ms) to create sequential charge-reduced precursor ion (CRI). The charges of the fragments and their sequences were determined with corresponding CRI. The product ions can be selected for subsequent CID (termed CIDn) combined with ETnoD for further sequence identification and validation. We refer to this method as CIDn/ETnoD. The use of a multistage CID activation (CIDn) and ETnoD protocol has been applied to several intact proteins to obtain multiple sequence identifications.
摘要：
在过去的几十年里，蛋白质组学已经变得越来越重要和被广泛讨论的话题。完整蛋白质的鉴定仍然是该领域的主要焦点。虽然大多数完整的蛋白质使用高分辨率质谱分析，通过低分辨率质谱鉴定它们继续带来挑战。在我们的研究中,我们研究了使用碰撞诱导解离（CID）和无解离电子转移（ETnoD）鉴定各种完整蛋白质的能力。用肌红蛋白作为我们的测试蛋白，用CID产生稳定的产物离子，用ETnoD鉴定产物离子的身份。ETnoD使用较短的激活时间(AcT，5ms）以创建顺序的电荷减少的前体离子（CRI）。用相应的CRI测定片段及其序列的电荷。可以选择产物离子用于与ETnoD组合的后续CID(termedCIDn),以用于进一步的序列识别和验证。我们将此方法称为CIDn/ETnod。多阶段CID激活（CIDn）和ETnoD方案的使用已应用于几种完整的蛋白质，以获得多个序列鉴定。
