High-throughput sequencing of peptoids and peptide-peptoid hybrids by partial edman degradation and mass spectrometry.
High-throughput sequencing of peptoids and peptide-peptoid hybrids by partial edman degradation and mass spectrometry.
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DOI：
10.1021/cc8001734
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发表时间：
2009-03-09
影响因子：
--
通讯作者：
Pei, Dehua
中科院分区：
文献类型：
--
作者：
Thakkar, Amit;Cohen, Allison S.;Connolly, Michael D.;Zuckermann, Ronald N.;Pei, Dehua
本文提出了一种从一珠一化合物组合文库中筛选类肽[寡（N-取代甘氨酸）]和肽-类肽杂合体的快速序列测定方法。在该方法中，通过用异硫氰酸苯酯（PITC）和9-芴基甲基氯甲酸酯（Fmoc-Cl）的1：3（mol/mol）混合物处理，使携带独特类肽（或肽-类肽）序列的珠粒经历多个循环的部分埃德曼降解（PED），以产生每种树脂结合的类肽的一系列N-末端截短产物。PED后，通过哌啶处理从N-末端和任何反应的侧链除去Fmoc基团。通过基质辅助激光解吸电离（MALDI）质谱法分析全长类肽及其截短产物的所得混合物，以揭示全长类肽的序列。该方法稍加修改，也可用于肽-类肽杂交体的序列测定。这种快速、高通量、灵敏、廉价的测序方法将极大地扩展组合类肽文库在生物医学和材料研究中的应用。
A method for the rapid sequence determination of peptoids [oligo(N-substituted glycines)] and peptide-peptoids hybrids selected from one-bead-one-compound combinatorial libraries has been developed. In this method, beads carrying unique peptoid (or peptide-peptoid) sequences were subjected to multiple cycles of partial Edman degradation (PED) by treatment with a 1:3 (mol/mol) mixture of phenyl isothiocyanate (PITC) and 9-fluorenylmethyl chloroformate (Fmoc-Cl) to generate a series of N-terminal truncation products for each resin-bound peptoid. After PED, the Fmoc group was removed from the N-terminus and any reacted side chains via piperidine treatment. The resulting mixture of the full-length peptoid and its truncation products was analyzed by matrix-assisted laser desorption ionization (MALDI) mass spectrometry, to reveal the sequence of the full-length peptoid. With a slight modification, the method was also effective in the sequence determination of peptide-peptoid hybrids. This rapid, high-throughput, sensitive, and inexpensive sequencing method should greatly expand the utility of combinatorial peptoid libraries in biomedical and materials research.
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作者：
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