A single-domain protein catenane of dihydrofolate reductase  

作　　者：Jing Fang Tianzuo Li Jiyeon Lee Dahye Im Lianjie Xu Yajie Liu Jongcheol Seo Wen-Bin Zhang 

机构地区：[1]Beijing National Laboratory for Molecular Sciences,Key Laboratory of Polymer Chemistry&Physics of Ministry of Education,Center for Soft Matter Science and Engineering,College of Chemistry and Molecular Engineering,Peking University,Beijing 100871,China [2]Department of Chemistry,Pohang University of Science and Technology(POSTECH),Pohang 37673,Republic of Korea [3]Beijing Academy of Artificial Intelligence,Beijing 100084,China

出　　处：《National Science Review》2023年第11期123-132,共10页国家科学评论（英文版）

基　　金：supported by the National Key Research and Development Program of China(2020YFA0908100);the National Natural Science Foundation ofChina(21991132,21925102,92056118,22101010,22201016 and 22201017);the Beijing National Laboratory for Molecular Sciences(BNLMS-CXXM202006);the National Research Foundation of Korea(2019R1C1C1008414 and 2020R1A5A1019141)。

摘　　要：A single-domain protein catenane refers to two mechanically interlocked polypeptide rings that fold synergistically into a compact and integrated structure,which is extremely rare in nature.Here,we report a single-domain protein catenane of dihydrofolate reductase(cat-DHFR).This design was achieved by rewiring the connectivity between secondary motifs to introduce artificial entanglement and synthesis was readily accomplished through a series of programmed and streamlined post-translational processing events in cells without any additional in vitro reactions.The target molecule contained few exogenous motifs and was thoroughly characterized using a combination of ultra-performance liquid chromatography–mass spectrometry,sodium dodecyl sulfate–polyacrylamide gel electrophoresis,protease cleavage experiments and ion mobility spectrometry–mass spectrometry.Compared with the linear control,cat-DHFR retained its catalytic capability and exhibited enhanced stability against thermal or chemical denaturation due to conformational restriction.These results suggest that linear proteins may be converted into their concatenated single-domain counterparts with almost identical chemical compositions,well-preserved functions and elevated stabilities,representing an entirely new horizon in protein science.

关 键 词：cellular synthesis CATENANE protein domain chemical topology DHFR 

分 类 号：Q75[生物学—分子生物学]