New method of peptide cleavage based on Edman degradation.
New method of peptide cleavage based on Edman degradation.
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DOI：
10.1007/s11030-013-9453-y
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发表时间：
2013-08
影响因子：
3.8
通讯作者：
Szewczuk Z
中科院分区：
文献类型：
--
作者：
Bąchor R;Kluczyk A;Stefanowicz P;Szewczuk Z
提出了一种基于苯硫海因（PTH）生成的N-酰化肽的直接切割方法.该程序可应用于酸稳定树脂，如TentaGel HL-NH。我们设计了一个可切割的接头，由一个赖氨酸残基与-氨基封闭的Boc，而-氨基用于肽合成。在肽组装完成后，使用三氟乙酸除去肽侧链中的保护基团，从而也释放接头中赖氨酸的-氨基。然后与异硫氰酸苯酯反应，然后酸解，导致从树脂中有效释放肽作为稳定的PTH衍生物。此外，2-（4-氮杂-1-氮鎓双环[2.2.2]辛基铵）乙酰基形式的固定电荷标签的应用提高了电离效率并降低了检测限，允许在亚飞摩尔范围内对肽进行ESI-MS/MS测序。所提出的策略与一珠一化合物肽库合成期间的标准条件相容。所开发的策略在组合化学中的适用性得到证实，使用一个小的训练库-糜蛋白酶底物。本文的在线版本（doi：10.1007/s11030-013-9453-y）包含补充材料，可供授权用户使用。
A straightforward cleavage method for N- acylated peptides based on the phenylthiohydantoin (PTH) formation is presented. The procedure could be applied to acid-stable resins, such as TentaGel HL–NH. We designed a cleavable linker that consists of a lysine residue with the -amino group blocked by Boc, whereas the -amino group is used for peptide synthesis. After the peptide assembly is completed, the protecting groups in peptide side chains are removed using trifluoroacetic acid, thus liberating also the -amino group of the lysine in the linker. Then the reaction with phenyl isothiocyanate followed by acidolysis causes an efficient peptide release from the resin as a stable PTH derivative. Furthermore, the application of a fixed charge tag in the form of 2-(4-aza-1-azoniabicyclo[2.2.2]octylammonium)acetyl group increases ionization efficiency and reduces the detection limit, allowing ESI-MS/MS sequencing of peptides in the subfemtomolar range. The proposed strategy is compatible with standard conditions during one-bead-one-compound peptide library synthesis. The applicability of the developed strategy in combinatorial chemistry was confirmed using a small training library of -chymotrypsin substrates. The online version of this article (doi:10.1007/s11030-013-9453-y) contains supplementary material, which is available to authorized users.
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影响因子：
15
作者：
Cheung, YW;Abell, C;Balasubramanian, S
通讯作者：
Balasubramanian, S
影响因子：
3.8
作者：
Porco, JA;Deegan, T;Wright, P
通讯作者：
Wright, P
影响因子：
2.1
作者：
Cydzik, Marzena;Rudowska, Magdalena;Szewczuk, Zbigniew
通讯作者：
Szewczuk, Zbigniew
影响因子：
2.9
作者：
Liu, RW;Lam, KS
通讯作者：
Lam, KS
影响因子：
--
作者：
Gerona-Navarro, Guillermo;Gonzalez-Muniz, Rosario;Royo, Miriam
通讯作者：
Royo, Miriam