Facile analysis and sequencing of linear and branched peptide boronic acids by MALDI mass spectrometry.
Facile analysis and sequencing of linear and branched peptide boronic acids by MALDI mass spectrometry.
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DOI：
10.1021/ac2002565
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发表时间：
2011-05-01
影响因子：
7.4
通讯作者：
Santos, Webster L.
中科院分区：
文献类型：
--
作者：
Crumpton, Jason B.;Zhang, Wenyu;Santos, Webster L.
人们对含有硼酸部分的多肽越来越感兴趣，因为它们具有治疗/诊断多种疾病的潜力，如癌症。多肽硼酸的用途可以通过获得大量的文库来扩大，这些文库可以快速和经济地去卷积。遗憾的是，目前使用质谱学的检测方法费时费力，而且会受到硼酸三聚的干扰，这需要耗时地分析脱水产物。当肽序列未知时，这些问题会加剧，如从头测序，特别是当存在多个硼酸部分时。因此，寻找一种快速、可靠、简便的多肽鉴定方法是非常重要的。在这里，我们报告了用基质辅助激光解吸/电离质谱仪(MALDI-MS)鉴定和测序含有多达五个硼酸基团的线状和支链多肽硼酸。采用制备频那醇硼酸酯的方法对多肽进行高效的MALDI分析。此外，还建立了一种以2，5-二羟基苯甲酸(DHB)为基质和衍生化试剂的多肽硼酸检测方法，实现了方便、原位、在线的酯化反应。最后，我们证明了可以用MALDI-MSMS分析来自单个小球的DHB修饰的多肽硼酸，验证了我们的方法用于支链多肽硼酸文库的鉴定和测序。
Interest in peptides incorporating boronic acid moieties is increasing due to their potential as therapeutics/diagnostics for a variety of diseases such as cancer. The utility of peptide boronic acids may be expanded with access to vast libraries that can be deconvoluted rapidly and economically. Unfortunately, current detection protocols using mass spectrometry are laborious and confounded by boronic acid trimerization, which requires time consuming analysis of dehydration products. These issues are exacerbated when the peptide sequence is unknown, as with de novo sequencing, and especially when multiple boronic acid moieties are present. Thus, a rapid, reliable and simple method for peptide identification is of utmost importance. Herein, we report the identification and sequencing of linear and branched peptide boronic acids containing up to five boronic acid groups by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Protocols for preparation of pinacol boronic esters were adapted for efficient MALDI analysis of peptides. Additionally, a novel peptide boronic acid detection strategy was developed in which 2,5-dihydroxybenzoic acid (DHB) served as both matrix and derivatizing agent in a convenient, in situ, on-plate esterification. Finally, we demonstrate that DHB-modified peptide boronic acids from a single bead can be analyzed by MALDI-MSMS analysis, validating our approach for the identification and sequencing of branched peptide boronic acid libraries.
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影响因子：
2.1
作者：
Hoeg-Jensen, T;Ridderberg, S;Markussen, J
通讯作者：
Markussen, J
影响因子：
--
作者：
Bryson, David I.;Zhang, Wenyu;Santos, Webster L.
通讯作者：
Santos, Webster L.
影响因子：
1.1
作者：
Manku, Sukhdev;Hall, Dennis G.
通讯作者：
Hall, Dennis G.
影响因子：
3.5
作者：
Wienand, A;Ehrhardt, C;Tapparelli, C
通讯作者：
Tapparelli, C
影响因子：
4.9
作者：
Arimori, S;Bell, ML;James, TD
通讯作者：
James, TD