SnoopLigase-Mediated Peptide-Peptide Conjugation and Purification.

SnoopLigase-Mediated Peptide-Peptide Conjugation and Purification.
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SnoopLigase 介导的肽-肽缀合和纯化。

DOI:
10.1007/978-1-0716-0928-6_2
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发表时间:
2021
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Buldun CM
Buldun CM
中科院分区:
--
文献类型:
--
作者:
Buldun CM

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将不同蛋白质连接在一起的共价键可以增强一系列应用的功能。我们已经开发了SnoopLigase肽-肽缀合方法,以通过异肽键容易且特异性地连接与肽SnoopTagJr或DogTag融合的蛋白质。SnoopLigase偶联已用于增强酶弹性和抗原寡聚化以增强疫苗效力。缀合后,SnoopLigase和未反应的底物可以通过固相固定SnoopLigase来除去,产生纯化的蛋白质-蛋白质缀合物。在这里,我们描述了设计标签融合蛋白,SnoopLigase纯化,和连接的SnoopTagJr和DogTag的程序。我们进一步定义了纯化连接产物和定量连接成功的步骤。
Covalently linking together different proteins can enhance functionality for a range of applications. We have developed the SnoopLigase peptide–peptide conjugation method to easily and specifically link proteins fused to the peptides SnoopTagJr or DogTag via an isopeptide bond. SnoopLigase conjugation has been applied for enhancing enzyme resilience and for antigen oligomerization to enhance vaccine efficacy. Following conjugation, SnoopLigase and unreacted substrates can be removed by solid-phase immobilization of SnoopLigase, yielding purified protein–protein conjugates. Here, we describe procedures for designing tag-fused proteins, SnoopLigase purification, and ligation of SnoopTagJr and DogTag. We further define steps for the purification of the ligated product and quantification of ligation success.
DOI: 10.1186/s12915-017-0440-0
发表时间: 2017-10-30
期刊: BMC biology
影响因子: 5.4
作者:
Oesterle S;Roberts TM;Widmer LA;Mustafa H;Panke S;Billerbeck S
通讯作者: Billerbeck S
合成生物工程催化肽结构单元之间牢不可破的连接
DOI: --
发表时间: 2017
期刊:
影响因子: --
作者:
Can M Buldun
通讯作者: Can M Buldun
DOI: 10.1021/jacs.7b13237
发表时间: 2018-02-28
影响因子: 15
作者:
Buldun, Can M.;Jean, Jisoo X.;Howarth, Mark
通讯作者: Howarth, Mark