Easy amino acid sequencing of sulfonated peptides using post-source decay on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer equipped with a variable voltage reflector
Easy amino acid sequencing of sulfonated peptides using post-source decay on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer equipped with a variable voltage reflector
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DOI：
10.1002/rcm.805
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发表时间：
2002-01-01
影响因子：
2
通讯作者：
Bhikhabhai, R
中科院分区：
文献类型：
--
作者：
Hellman, U;Bhikhabhai, R
使用改进的化学方法在n端用磺酸基团标记色氨酸肽。衍生化在普通水缓冲液中对吸附在ZipTip(TM) C-18上的肽进行，从而允许同时对样品进行脱盐/浓缩。当只考虑精氨酸终止肽时，从吸附到ZipTip直到MALDI-PSD分析的过程大约需要10分钟，多个样品可以并行处理。由此产生的改进后源衰变（PSD）碎片产生仅含y离子的光谱。比较了未分化和衍生化合成肽的PSD氨基酸序列。从胰蛋白酶凝胶酶切中离子选择衍生肽获得的序列信息中，正确地鉴定出了一种难以从不明确的肽质量指纹分析中分析的蛋白质。该方法还应用于天然肽和合成肽中磷酸化Ser和Tyr残基的鉴定和定位。版权所有:John Wiley Sons, Ltd。
Tryptic peptides were labeled with sulfonic acid groups at the N-termini using an improved chemistry. The derivatization was performed in common aqueous buffers on peptides adsorbed onto a ZipTip(TM) C-18, thus allowing simultaneous desalting/concentration of the sample. When only Arg-terminating peptides were considered, the procedure from adsorption onto the ZipTip until analysis by MALDI-PSD took about 10 min and several samples could be worked on in parallel. The resulting improved post-source decay (PSD) fragmentation produced spectra containing only y-ions. PSD amino acid sequencing of underivatized and derivatized synthetic peptides was compared. From the sequence information obtained from derivatized peptides isolated by ion selection from tryptic in-gel digests, a protein was correctly identified which was difficult to analyze from an unclear peptide mass fingerprint analysis. The method was also applied to the identification and localization of phosphorylated Ser and Tyr residues in native and synthetic peptides. Copyright (C) 2002 John Wiley Sons, Ltd.