题目：Metalloprotein design using genetic code expansion
Metalloprotein design using genetic code expansion
Summary: One aim in our laboratory is to use small, soluble protein scaffold, and the genetic incorporation of unnatural amino acid to design easy-to-characterize, easy-to-produce, and easy-to-optimize metalloenzymes which catalyze these important reactions with equal or greater efficiency/selectivity than that of the natural systems. Through the genetic incorporation of the Tyr-His ligand and CuB site into myoglobin, we recapitulated important features of HCO into this small soluble protein, which exhibits selective O2 reduction activity while generating less than 6% ROS, at more than 1000 turnovers. These results support that Tyr-His crosslink is indeed important for HCO function, and creates the exciting opportunity to rapidly evolve better HCO model proteins to achieve higher activity and selectivity, which may be suitable as alternatives to precious metal catalyst in fuel cells.
Another aspect of our ongoing research is the development of new methods for precise attachment of functional metal complexes on biomolecules, which is an important strategy for metalloprotein design. Bioorthogonal chemical reactions together with genetic code expansion technique have provided exciting new means for protein labeling in living cells. The main advantages of photoclick reaction are its fast rate (up to 50 M-1S-1), and that it has no need for toxic copper catalyst.
Recent representative publications (as a corresponding author)
1. Li, F. H.; Shi P.; Li J. S.;Yang F.; Wang T. Y.; Zhang W.; Gao F.; Ding W.; Li D.; Li J.; Xiong Y.; Sun J. P.; Gong W. M.; Tian C. L.; Wang J. Y., “A Genetically Encoded 19F NMR Sensor for Tyrosine Phosphorylation ” Angew. Chem. Intl. Ed. 2013, 52, 3958-62.
2. Zhou Q.; Hu M. R.; Zhang W.; Jiang, L.; Perrett S.; Zhou J.; Wang J. Y., “Probing the Function of the Tyr-Cys Crosslink in Metalloenzymes through the Genetic Incorporation of 3-Methylthiotyrosine” Angew. Chem. Intl. Ed. 2013, 52, 1203-7.
3. Liu, X. H.; Yu, Y.; Hu, C.; Zhang, W.; Lu, Y.; Wang, J. Y, “Significant Increase of Oxidase Activity through the Genetic Incorporation of a Tyrosine-Histidine Cross-Link in a Myoglobin Model of Heme-Copper Oxidase” Angew. Chem. Intl. Ed. 2012, 51 , 4312-6. (News of the Week, Chemical and Engineering News)
4.. Liu, X. H.; Li, J. S.; Dong, J. S.; Hu , C.; Gong, W. M.; Wang, J. Y., “Genetic Incorporation of a Metal Chelating Amino Acid as a Probe for Protein Electron Transfer” Angew. Chem. Intl. Ed. 2012, 51, 10261-5. (Very Important Paper and Cover Article)
5. Yu, Z.; Pan, Y. C.; Wang. Z. Y.; Wang, J. Y.; Lin. Q., “Genetically Encoded Cyclopropene Directs Rapid, Photoclick Chemistry- Mediated Protein Labeling in Mammalian Cells” Angew. Chem. Intl. Ed. 2012, 51，10600-4.
6. Wang, J. Y.; Zhang, W.; Song, W. J.; Wang, Y. Z.; Yu, Z. P.; Li, J. S.; Wu, M. H.; Wang, L.; Zang, J. Y.; Lin, Q., “A Biosynthetic Route to Photoclick Chemistry on Proteins” J. Am. Chem. Soc. 2010, 132, 14812-8.
7. Liu, X. H.; Li J. S.; Hu C.; Zhang W.; Hu M. R.; Zhou J.; Wang J. Y., “Significant Expansion of the Fluorescent Protein Chromophore through the Genetic Incorporation of an Metal-chelating Unnatural Amino Acid” Angew. Chem. Intl. Ed. 2013, 52, 4805-9
8. Li, F. H.; Zhang, H.; Sun, Y.; Pan, Y. C.; Zhou, J. Z.; Wang, J. Y., “Expanding the Genetic Code for Photoclick Chemistry in E. coli, Mammalian Cells and A. thaliana” Angew. Chem. Intl. Ed. 2013 DOI: 10.1002/anie. 201303477
C.V. of Jiangyun Wang
Jiangyun Wang holds a diploma degree in Physical Chemistry from the University of Science and Technology of China. He finished his PhD under the supervision of Kenneth Suslick at the University of Illinois at Urbana-Champaign in 2003. Thereafter, he worked as a Research Fellow in the Peter Schultz Group of the Scripps Research Institute in San Diego, California. Since January 2008 he is head of the Institute of Biophysics Group for Chemical Biology. His present research interests includes:
1. Genetic code expansion to facilitate protein labeling and engineering in various host organisms, including bacteria cyanobacteria, mammals and plants.
2. Development of new bio-orthogonal click chemical reactions to facilitate protein and RNA labeling, as wells as super-resolution imaging in living cells.
3. Design of metalloproteins, which includes cytochrome c oxidase, nitrogenase, photosystem II.