a man-made atp-binding protein evolved independent of nature causes abnormal growth in bacterial cells人造磷酸腺苷蛋白进化独立于自然导致细菌细胞异常生长.pdfVIP

A Man-Made ATP-Binding Protein Evolved Independent of Nature Causes Abnormal Growth in Bacterial Cells 1,4 2¤ ´ 1 3 1,5 Joshua M. Stomel , James W. Wilson , Megan A. Leon , Phillip Stafford , John C. Chaput * 1 Center for BioOptical Nanotechnology, Arizona State University, Tempe, Arizona, United States of America, 2 Center for Infectious Disease and Vaccinology, Arizona State University, Tempe, Arizona, United States of America, 3 Center for Innovations in Medicine, The Biodesign Institute, Arizona State University, Tempe, Arizona, United States of America, 4 School of Life Sciences, Arizona State University, Tempe, Arizona, United States of America, 5 Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona, United States of America Abstract Recent advances in de novo protein evolution have made it possible to create synthetic proteins from unbiased libraries that fold into stable tertiary structures with predefined functions. However, it is not known whether such proteins will be functional when expressed inside living cells or how a host organism would respond to an encounter with a non-biological protein. Here, we examine the physiology and morphology of Escherichia coli cells engineered to express a synthetic ATP- binding protein evolved entirely from non-biological origins. We show that this man-made protein disrupts the normal energetic balance of the cell by altering the levels of intracellular ATP. This disruption cascades into a series of events that ultimately limit reproductive competency by inhibiting cell division. We now describe a detailed investigation into the synthetic biology of this man-made protein in a liv