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适用于固相微量测序的肽片段化。N-溴代琥珀酰亚胺和BNPS-粪臭素(3-溴-3-甲基-2-[(2-硝基苯基)硫代]-3H-吲哚)的应用。

Peptide fragmentation suitable for solid-phase microsequencing. Use of N-bromosuccinimide and BNPS-skatole (3-bromo-3-methyl-2-[(2-nitrophenyl)thio]-3H-indole).

作者信息

Hunziker P E, Hughes G J, Wilson K J

出版信息

Biochem J. 1980 May 1;187(2):515-9. doi: 10.1042/bj1870515.

Abstract

BNPS-skatole (3-bromo-3-methyl-2-(2-nitrophenyl)thiol-3H-indole) and N-bromosuccinimide have been used to specifically cleave at peptide bonds after amino acids with available C-gamma=C-delta double bonds, i.e. tryptophan, tyrosine and histidine. The resulting C-terminal lactones conveniently attach to amino-glass supports for sequencing with DABITC (4-NN-dimethylaminoazobenzene 4'-isothiocyanate). Also, peptides having such C-termini, i.e. from a chymotryptic digest, can be readily made to react with these reagents and thus be easily attached and sequenced by solid-phase methods.

摘要

BNPS-粪臭素(3-溴-3-甲基-2-(2-硝基苯基)硫醇-3H-吲哚)和N-溴代琥珀酰亚胺已被用于在具有可利用的C-γ=C-δ双键的氨基酸(即色氨酸、酪氨酸和组氨酸)之后的肽键处进行特异性切割。所得的C-末端内酯可方便地连接到氨基玻璃支持物上,以便用DABITC(4-N,N-二甲基氨基偶氮苯4'-异硫氰酸酯)进行测序。此外,具有此类C-末端的肽,即来自胰凝乳蛋白酶消化产物的肽,可很容易地与这些试剂反应,从而通过固相方法轻松连接并测序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d28a/1161818/b549ea4ca67b/biochemj00425-0230-a.jpg

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