Unlike small molecules,the topological complexity of macromolecules remains largely unexplored due to the huge synthetic challenge.Herein,we report the development of orthogonal active templates for concise and select...Unlike small molecules,the topological complexity of macromolecules remains largely unexplored due to the huge synthetic challenge.Herein,we report the development of orthogonal active templates for concise and selective synthesis of protein[n]heterocatenanes toward protein olympiadanes.An active template(AT-Snoop)was first developed based on the isopeptide-bond-forming RrgA domain with comparable efficiency and excellent orthogonality to the previously reported active template(AT-Spy)based on the CnaB2 domain.Their combination facilitated the selective synthesis of protein[n]catenanes from multiple components in one step and the resulting structures were verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis,size exclusion chromatography,liquid chromatography-mass spectrometry,and proteolytic digestion experiments.The results offered a promising solution to tackling the daunting challenge of precision synthesis of protein olympiadane with five distinct ring components.Not only did the success provide new tools for proteintopology engin eering but alsospurred and fueled the future exploitation of topology-related functional benefits in protein science.展开更多
基金the National Natural Science Foundation of China(grant nos.21991132,21925102,92056118,22101010,22201017,and 22201016)the National Key R&D Program of China(grant no.2020YFA0908100)+1 种基金Beijing National Laboratory for Molecular Sciences(grant no.BNLMS-CXXM-202006)supported by the National Center for Protein Sciences at Peking University.
文摘Unlike small molecules,the topological complexity of macromolecules remains largely unexplored due to the huge synthetic challenge.Herein,we report the development of orthogonal active templates for concise and selective synthesis of protein[n]heterocatenanes toward protein olympiadanes.An active template(AT-Snoop)was first developed based on the isopeptide-bond-forming RrgA domain with comparable efficiency and excellent orthogonality to the previously reported active template(AT-Spy)based on the CnaB2 domain.Their combination facilitated the selective synthesis of protein[n]catenanes from multiple components in one step and the resulting structures were verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis,size exclusion chromatography,liquid chromatography-mass spectrometry,and proteolytic digestion experiments.The results offered a promising solution to tackling the daunting challenge of precision synthesis of protein olympiadane with five distinct ring components.Not only did the success provide new tools for proteintopology engin eering but alsospurred and fueled the future exploitation of topology-related functional benefits in protein science.
