Mass spectrometry evidence for cisplatin as a protein cross-linking reagent.
Mass spectrometry evidence for cisplatin as a protein cross-linking reagent.
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DOI：
10.1021/ac200861k
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发表时间：
2011-07-01
影响因子：
7.4
通讯作者：
O'Connor, Peter B.
中科院分区：
文献类型：
--
作者：
Li, Huilin;Zhao, Yao;Phillips, Hazel I. A.;Qi, Yulin;Lin, Tzu-Yung;Sadler, Peter J.;O'Connor, Peter B.
顺铂是一种有效的抗癌药物，其功能是通过使相邻的DNA鸟嘌呤残基交联来发挥作用。然而，在注射后一天内，血浆中65~98%的铂是蛋白质结合的。人们普遍认为顺铂与蛋氨酸和组氨酸残基结合，但经常被低估的是，顺铂中的铂带有2+电荷，可以形成多达四个键。因此，它有可能作为交联剂发挥作用。用标准多肽--16.8 kDa蛋白钙调蛋白(CaM)，用傅里叶变换离子回旋共振(FTICR)质谱仪研究了顺铂的交联能力，但对kDa蛋白血红蛋白的交联作用不明显。FTICR MS的高分辨率和高质量准确度以及碰撞激活解离(CAD)和电子捕获解离(ECD)提供的大分子多肽的高度碎裂被证明是表征交联点的有价值的手段。顺铂不同于目前的交联剂，它针对新的官能团、硫醚和咪唑基，与现有的交联剂相辅相成。此外，铂(II)本身具有两个正电荷，这增强了对交联产物的检测。较高的电荷态不仅促进了纯度较低的交联物的检测，而且导致了更全面的MS/MS裂解，有助于修饰位点的分配。此外，铂的独特同位素模式通过质谱学标记了交联物和修饰部位。
Cisplatin is a potent anti-cancer drug, which functions by cross-linking adjacent DNA guanine residues. However within one day of injection, 65~98% of the platinum in the blood plasma is protein-bound. It is generally accepted that cisplatin binds to methionine and histidine residues, but what is often underappreciated is that platinum from cisplatin has a 2+ charge and can form up to four bonds. Thus, it has the potential to function as a cross-linker. In this report, the cross-linking ability of cisplatin is demonstrated by Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) with the use of standard peptides, the 16.8 kDa protein calmodulin (CaM), but was unsuccessful for the 64 kDa protein hemoglobin. The high resolution and mass accuracy of FTICR MS along with the high degree of fragmentation of large peptides afforded by collisionally activated dissociation (CAD) and electron capture dissociation (ECD) are shown to be a valuable means of characterizing cross-linking sites. Cisplatin is different from current cross-linking reagents by targeting new functional groups, thioethers, and imidazoles groups, which provides complementarity with existing cross-linkers. In addition, platinum(II) inherently has two positive charges which enhance the detection of cross-linked products. Higher charge states not only promote the detection of cross-linking products with less purification, but result in more comprehensive MS/MS fragmentation and can assist the assignment of modification sites. Moreover, the unique isotopic pattern of platinum flags cross-linking products and modification sites by mass spectrometry.
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影响因子：
4.6
作者：
Hartinger, Christian G.;Tsybin, Yury O.;Dyson, Paul J.
通讯作者：
Dyson, Paul J.
DOI：
10.1002/oms.1210260527
发表时间：
1991-05-01
期刊：
ORGANIC MASS SPECTROMETRY
影响因子：
--
作者：
CARAVATTI, P;ALLEMANN, M
通讯作者：
ALLEMANN, M
影响因子：
7.3
作者：
Knipp, Markus;Karotki, Andrei V.;Vasak, Milan
通讯作者：
Vasak, Milan
影响因子：
4.3
作者：
Lau, JKC;Deubel, DV
通讯作者：
Deubel, DV
影响因子：
4.6
作者：
Ienco, Andrea;Caporali, Maria;Mealli, Carlo
通讯作者：
Mealli, Carlo