Silva Maria J S A, Faustino Hélio, Coelho Jaime A S, Pinto Maria V, Fernandes Adelaide, Compañón Ismael, Corzana Francisco, Gasser Gilles, Gois Pedro M P
Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Lisbon, Portugal.
Chimie ParisTech, PSL University, CNRS, Institute of Chemistry for Life and Health Sciences, Laboratory for Inorganic Chemical Biology, 75005, Paris, France.
Angew Chem Int Ed Engl. 2021 May 3;60(19):10850-10857. doi: 10.1002/anie.202016936. Epub 2021 Apr 1.
Widely used reagents in the peptide functionalization toolbox, Michael acceptors and N-hydroxysuccinimide (NHS) activated esters, are combined in NHS-activated acrylamides for efficient chemoselective amino-sulfhydryl stapling on native peptides and proteins. NHS-activated acrylamides allow for a fast functionalization of N-terminal cysteines (k =1.54±0.18×10 M s ) under dilute aqueous conditions, enabling selectivity over other nucleophilic amino acids. Additionally, the versatility of these new bioconjugation handles was demonstrated in the cross-linking of in-chain or C-terminal cysteines with nearby lysine residues. NHS-activated acrylamides are compatible with the use of other cysteine selective reagents, allowing for orthogonal dual-modifications. This strategy was successfully applied to the late-stage functionalization of peptides and proteins with a PEG unit, fluorescent probe, and cytotoxic agent. The level of molecular control offered by NHS-activated acrylamides is expected to promote amino-sulfhydryl stapling technology as a powerful strategy to design functional bioconjugates.
肽功能化工具箱中广泛使用的试剂——迈克尔受体和N-羟基琥珀酰亚胺（NHS）活化酯，被组合成NHS活化的丙烯酰胺，用于在天然肽和蛋白质上进行高效的化学选择性氨基-巯基钉合。NHS活化的丙烯酰胺能够在稀水溶液条件下快速官能化N端半胱氨酸（k = 1.54±0.18×10 M s），从而实现对其他亲核氨基酸的选择性。此外，这些新型生物共轭基团在链内或C端半胱氨酸与附近赖氨酸残基的交联中展现出了多功能性。NHS活化的丙烯酰胺与其他半胱氨酸选择性试剂兼容，允许进行正交双修饰。该策略已成功应用于用聚乙二醇单元、荧光探针和细胞毒性剂对肽和蛋白质进行后期功能化。预计NHS活化的丙烯酰胺所提供的分子控制水平将推动氨基-巯基钉合技术成为设计功能性生物共轭物的有力策略。
