Automatic Edman microsequencing of peptides containing multiple unnatural amino acids
Automatic Edman microsequencing of peptides containing multiple unnatural amino acids
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DOI：
10.1006/abio.2001.5172
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发表时间：
2001-08-01
影响因子：
2.9
通讯作者：
Lam, KS
中科院分区：
文献类型：
--
作者：
Liu, RW;Lam, KS
现在常规使用自动Edman微测序来确定含有天然氨基酸的肽或蛋白质的一级结构;然而，容易测序含有非天然氨基酸的肽的能力的缺陷仍然存在。随着合成肽化学、组合化学和大量可商购的非天然氨基酸的出现，需要对含有许多非天然氨基酸的短肽进行有效且准确的结构测定。在这项研究中，选择了35种市售的非天然氨基酸，以确定它们在ABI蛋白测序仪上的洗脱曲线。使用稍微修改的梯度程序，这35种PTH氨基酸中的19种可以容易地分辨，并在低皮摩尔水平下与常见的PTH氨基酸区分开。这些非天然氨基酸与20种天然氨基酸结合可用作构建肽文库的结构单元，并且从这些文库分离的肽珠可容易地进行微测序。为了证明这一点，我们合成了一个简单的三肽“一珠一化合物”的组合库，含有14个非天然和19个天然氨基酸，并筛选该库的链霉亲和素结合配体。分离的肽珠的微测序揭示了新的基序Bpa-Phe（4-X）-Aib，其中X = H、OH和CH 3。(C)北京：科学出版社.
It is now routine using automatic Edman microsequencing to determine the primary structure of peptides or proteins containing natural amino acids; however, a deficiency in the ability to readily sequence peptides containing unnatural amino acids remains. With the advent of synthetic peptide chemistry, combinatorial chemistry, and the large number of commercially available unnatural amino acids, there is a need for efficient and accurate structure determination of short peptides containing many unnatural amino acids. In this study, 35 commercially available a-un-natural amino acids were selected to determine their elution profile on an ABI protein sequencer. Using a slightly modified gradient program, 19 of these 35 PTH amino acids can be readily resolved and distinguished from common PTH amino acids at low picomole levels. These unnatural amino acids in conjunction with the 20 natural amino acids can be used as building blocks to construct peptide libraries, and peptide beads isolated from these libraries can be readily microsequenced. To demonstrate this, we synthesized a simple tripeptide "one-bead one-compound" combinatorial library containing 14 unnatural and 19 natural amino acids and screened this library for streptavidin-binding ligands. Microsequencing of the isolated peptide-beads revealed the novel motif Bpa-Phe(4-X)-Aib, wherein X = H, OH, and CH3. (C) 2001 Academic Press.