Mass Spectrometry Cleavable Strategy for Identification and Differentiation of Prenylated Peptides
Mass Spectrometry Cleavable Strategy for Identification and Differentiation of Prenylated Peptides
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DOI：
10.1021/ac503794s
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发表时间：
2015-02-17
影响因子：
7.4
通讯作者：
Chowdhury, Saiful M.
中科院分区：
文献类型：
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作者：
Bhawal, Ruchika P.;Sadananda, Sandhya C.;Chowdhury, Saiful M.
蛋白质的异戊烯化（法尼基化和香叶基香叶基化）与多种人类癌症有关，例如胰腺癌、结肠癌和急性骨髓性白血病，以及哈钦森-吉尔福德早老综合征（HGPS），这是一种与儿童过早衰老相关的遗传性疾病。目前的生物化学方法在体内或体外识别和区分大规模异戊烯化方面不是很有效。有有限的方法可用于大规模检测异戊二烯化的蛋白质，使用质谱，目前没有可用的方法，可以区分法尼基化和香叶基香叶基修饰在一个单一的实验设置。在这项研究中，一个简单的和新的方法检测和区分大规模的异戊二烯化肽使用质谱裂解的方法。该方法利用异戊二烯基上的简单化学和气相中亚砜基的可裂解性质，在串联质谱事件期间产生特征质谱。从修饰的异戊二烯化肽损失的特征质量区分异戊二烯化的类型。我们还在蛋白质的异戊烯化位点引入了环氧基团，使其更具亲水性，并可从复杂样品中富集。环氧基团的稳定性也在液相色谱质谱（LCMS）条件下进行了研究。该方法的概念验证是使用模拟蛋白质中的异戊二烯基序的异戊二烯化肽建立的。我们相信这种方法将在大规模研究中推进蛋白质中异戊二烯化类型的鉴定和区分，并将显著提高我们对癌症机制，癌症治疗和诊断的认识。
Prenylation of protein (farnesylation and geranylgeranylation) is involved in several human cancers, such as pancreatic, colon, and acute myeloid leukemia as well as Hutchinson-Gilford progeria syndrome (HGPS), a genetic disease that is associated with premature aging for children. Current biochemical methods are not very efficient in identifying and differentiating large-scale prenylations in vivo or in vitro. There are limited methods available for large-scale detection of prenylated proteins using mass spectrometry and no methods currently available which can distinguish farnesylation and geranylgeranylation modification in a single experimental setup. In this study, a simple and novel method for detection and distinction of large-scale prenylated peptides using mass spectrometry-cleavable approaches was developed. The method utilizes simple chemistry on the prenyl group and cleavable properties of a sulfoxide group in the gas phase to produce a signature mass spectrum during tandem mass spectrometric events. The characteristic masses lost from the modified prenylated peptides distinguished the types of prenylation. We also introduced epoxy groups in the prenylation sites of the proteins to make them more hydrophilic and enrichable from complex samples. Stability of the epoxide group was also studied under liquid chromatographymass spectrometry (LCMS) conditions. The proof-of-concept of this method was established using prenylated peptides which mimicked the prenyl motifs in the proteins. We believe this method will advance the identification and differentiation of the types of prenylation in proteins in large-scale studies and will improve significantly our knowledge of the mechanism of cancer, cancer treatments, and diagnosis.