Aspects of oligonucleotide and peptide sequencing with MALDI and electrospray mass spectrometry.
Aspects of oligonucleotide and peptide sequencing with MALDI and electrospray mass spectrometry.
复制标题
使用 MALDI 和电喷雾质谱法进行寡核苷酸和肽测序。
DOI：
10.1016/s0968-0896(98)00098-4
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发表时间：
1998
影响因子：
3.5
通讯作者：
Siuzdak,G
中科院分区：
文献类型：
--
作者：
Owens,DR;Bothner,B;Phung,Q;Harris,K;Siuzdak,G
随着基质辅助激光解吸/电离（MALDI）和电喷雾电离（ESI）技术的发展，生物聚合物质谱测序变得越来越重要。在这里，我们研究了顺序消化快速鉴定蛋白水解片段的使用，反过来强调了酶促MALDI阶梯测序和ESI串联质谱的一般用途。分析范围从2到50个残基的寡核苷酸，从7到44个残基的肽和病毒外壳蛋白。利用外切酶酶切法进行MALDI阶梯测序，产生离子均匀分布，并提供2-30个核酸残基长的寡核苷酸完整序列信息。仅获得了较长寡核苷酸的部分序列信息。c端肽阶梯测序通常提供4到7个氨基酸进入肽的信息。顺序消化，或内源性蛋白酶后外源性蛋白酶暴露，也成功地应用于胰蛋白酶消化病毒蛋白。与MALDI-MS分析相比，LCMS分析阶梯测序肽产生的信息较少，esi - ms2通常提供小寡核苷酸和肽的部分序列信息。总的来说，MALDI阶梯测序提供了比esi - ms2更广泛的生物聚合物的信息，并且相对简单易懂，特别是对于寡核苷酸。
Biopolymer sequencing with mass spectrometry has become increasingly important and accessible with the development of matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). Here we examine the use of sequential digestion for the rapid identification of proteolytic fragments, in turn highlighting the general utility of enzymatic MALDI ladder sequencing and ESI tandem mass spectrometry. Analyses were performed on oligonucleotides ranging in size from 2 to 50 residues, on peptides ranging in size from 7 to 44 residues and on viral coat proteins. MALDI ladder sequencing using exonuclease digestion generated a uniform distribution of ions and provided complete sequence information on the oligonucleotides 2–30 nucleic acid residues long. Only partial sequence information was obtained on the longer oligonucleotides. C-terminal peptide ladder sequencing typically provided information from 4 to 7 amino acids into the peptide. Sequential digestion, or endoprotease followed by exoprotease exposure, was also successfully applied to a trypsin digest of viral proteins. Analysis of ladder sequenced peptides by LCMS generated less information than in the MALDI-MS analysis and ESI-MS2normally provided partial sequence information on both the small oligonucleotides and peptides. In general, MALDI ladder sequencing offered information on a broader mass range of biopolymers than ESI-MS2and was relatively straightforward to interpret, especially for oligonucleotides.