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Cellular Synthesis and X-ray Crystal Structure of a Designed Protein Heterocatenane |
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| Authors: | Yajie Liu Zelin Duan Jing Fang Fan Zhang Prof Junyu Xiao Prof Wen-Bin Zhang |
| Institution: | 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 P. R. China
These authors contributed equally to the work.;2. State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, 100871 P. R. China These authors contributed equally to the work.;3. 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 P. R. China;4. State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, 100871 P. R. China |
| Abstract: | Herein, we report the biosynthesis of protein heterocatenanes using a programmed sequence of multiple post-translational processing events including intramolecular chain entanglement, in situ backbone cleavage, and spontaneous cyclization. The approach is general, autonomous, and can obviate the need for any additional enzymes. The catenane topology was convincingly proven using a combination of SDS-PAGE, LC-MS, size exclusion chromatography, controlled proteolytic digestion, and protein crystallography. The X-ray crystal structure clearly shows two mechanically interlocked protein rings with intact folded domains. It opens new avenues in the nascent field of protein-topology engineering. |
| Keywords: | catenanes inteins protein design protein engineering protein structures |