Site-Specific Reversible Protein and Peptide Modification: Transglutaminase-Catalyzed Glutamine Conjugation and Bioorthogonal Light-Mediated Removal.
Site-Specific Reversible Protein and Peptide Modification: Transglutaminase-Catalyzed Glutamine Conjugation and Bioorthogonal Light-Mediated Removal.
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DOI：
10.1021/acs.bioconjchem.9b00145
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发表时间：
2019-04
影响因子：
4.7
通讯作者：
Kevin R Moulton;A. Sadiki;Bilyana N Koleva;Lincoln J Ombelets;Tina H Tran;Shanshan Liu;Bryan Wang;Hongyan Chen;Emily Micheloni;P. Beuning;G. O'Doherty;Z. Zhou
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作者：
Kevin R Moulton;A. Sadiki;Bilyana N Koleva;Lincoln J Ombelets;Tina H Tran;Shanshan Liu;Bryan Wang;Hongyan Chen;Emily Micheloni;P. Beuning;G. O'Doherty;Z. Zhou
动态光开关赋予蛋白质的空间和时间控制是重要的操作和研究生物和非生物过程。然而，将这些开关安装到蛋白质特定位点的方法是有限的。在这里，我们描述了一种新的位点特异性方法来产生光去除蛋白缀合物。含胺的发色团（例如，古老的邻硝基苯基和较少探索的邻硝基苯基乙基）通过谷氨酰胺转氨酶（TGase, EC 2.3.2.13）在α-麦胶蛋白、LCMV和TAT肽以及β-酪蛋白和UmuD蛋白的谷氨酰胺侧链上转氨化。随后，光解再生了天然多肽和蛋白质。当这种修饰导致某些活性的减少或消除时，这个过程被称为笼化，就像大肠杆菌聚合酶管理蛋白UmuD的情况一样。重要的是，该方法简单、健壮且易于适应，例如，所有组件都是市售的。
Dynamic photoswitches in proteins that impart spatial and temporal control are important to manipulate and study biotic and abiotic processes. Nonetheless, approaches to install these switches into proteins site-specifically are limited. Herein we describe a novel site-specific method to generate photoremovable protein conjugates. Amine-containing chromophores (e.g., venerable o-nitrobenzyl and less-explored o-nitrophenylethyl groups) were incorporated via transamidation into a glutamine side chain of α-gliadin, LCMV, and TAT peptides, as well as β-casein and UmuD proteins by transglutaminase (TGase, EC 2.3.2.13). Subsequently, photolysis regenerated the native peptides and proteins. When this modification leads to the reduction or abolishment of certain activities, the process is referred to as caging, as in the case for E. coli polymerase manager protein UmuD. Importantly, this method is simple, robust, and easily adaptable, e.g., all components are commercially available.