Collision induced dissociation products of disulfide-bonded peptides: ions result from the cleavage of more than one bond.
Collision induced dissociation products of disulfide-bonded peptides: ions result from the cleavage of more than one bond.
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DOI：
10.1007/s13361-010-0064-x
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发表时间：
2011-03
影响因子：
3.2
通讯作者：
Desaire, Heather
中科院分区：
文献类型：
--
作者：
Clark, Daniel F.;Go, Eden P.;Toumi, Melinda L.;Desaire, Heather
二硫键是一种翻译后修饰（PTM），可以在重组表达、样品处理或样品纯化过程中被打乱或洗牌成非天然键。目前，由于各种样品要求和数据分析困难，二硫键的映射是困难的。促进这项困难工作的一个步骤是更好地理解二硫键肽在碰撞诱导解离（CID）过程中是如何分裂的。大多数自动分析算法都是基于这样的假设，即在二硫键结合的肽解离过程中观察到的产物离子的优势是由于一个肽键的断裂造成的，在本报告中，我们通过广泛分析几个二硫键结合的肽在QTOF仪器上受到CID时产生的产物离子来验证这一假设。我们发现最常见的产物离子类型之一是由两个肽键裂解或双重裂解产生的。我们发现，对于分析的一些二硫键肽，双裂解产物离子的数量超过了单裂解产物离子的数量。研究了电荷状态和前驱体离子大小的影响，以确定这些参数是否决定了形成双裂解产物离子的数量。在这组样品中发现，电荷状态或肽大小与分配为双裂的产物离子的部分之间没有很强的相关性。这些数据表明，这些离子可以解释在二硫键合肽的CID数据中检测到的许多产物离子。我们还展示了双裂解产物离子在具有多个半胱氨酸存在的肽上的效用。双裂解产物能够充分表征每种半胱氨酸的键合模式，而典型的单b/y裂解产物则不能。
Disulfide bonds are a posttranslational modification (PTM) that can be scrambled or shuffled to non-native bonds during recombinant expression, sample handling, or sample purification. Currently, mapping of disulfide bonds is difficult due, to various sample requirements and data analysis difficulties. One step towards facilitating this difficult work is developing a better understanding of how disulfide-bonded peptides fragment during Collision Induced Dissociation (CID). Most automated analysis algorithms function based on the assumption that the preponderance of product ions observed during the dissociation of disulfide-bonded peptides result from the cleavage of just one peptide bond, and in this report we tested that assumption by extensively analyzing the product ions generated when several disulfide-bonded peptides are subjected to CID on a QTOF instrument. We found that one of the most common types of product ions generated resulted from two peptide bond cleavages, or a double cleavage. We found that for several of the disulfide-bonded peptides analyzed, the number of double cleavage product ions outnumbered those of single cleavages. The influence of charge state and precursor ion size was investigated, to determine if those parameters dictated the amount of double cleavage product ions formed. It was found in this sample set that no strong correlation existed between the charge state or peptide size and the portion of product ions assigned as double cleavages. This data shows that these ions could account for many of the product ions detected in CID data of disulfide bonded peptides. We also showed the utility of double cleavage product ions on a peptide with multiple cysteines present. Double cleavage products were able to fully characterize the bonding pattern of each cysteine where typical single b/y cleavage products could not.
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影响因子：
4.4
作者：
Panchaud, Alexandre;Singh, Pragya;Shaffer, Scott A.;Goodlett, David R.
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影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
McLuckey, SA