| Autor: |
Markhard AL; Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114.; Broad Institute of MIT and Harvard, Cambridge, MA 02142.; Department of Systems Biology, Harvard Medical School, Boston, MA 02115., McCoy JG; Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114.; Broad Institute of MIT and Harvard, Cambridge, MA 02142.; Department of Systems Biology, Harvard Medical School, Boston, MA 02115., To TL; Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114.; Broad Institute of MIT and Harvard, Cambridge, MA 02142.; Department of Systems Biology, Harvard Medical School, Boston, MA 02115., Mootha VK; Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114.; Broad Institute of MIT and Harvard, Cambridge, MA 02142.; Department of Systems Biology, Harvard Medical School, Boston, MA 02115. |
| Abstrakt: |
Oxygen plays a key role in supporting life on our planet. It is particularly important in higher eukaryotes where it boosts bioenergetics as a thermodynamically favorable terminal electron acceptor and has important roles in cell signaling and development. Many human diseases stem from either insufficient or excessive oxygen. Despite its fundamental importance, we lack methods with which to manipulate the supply of oxygen with high spatiotemporal resolution in cells and in organisms. Here, we introduce a genetic system, SupplemeNtal Oxygen Released from ChLorite (SNORCL), for on-demand local generation of molecular oxygen in living cells, by harnessing prokaryotic chlorite O 2 -lyase (Cld) enzymes that convert chlorite (ClO 2 - ) into molecular oxygen (O 2 ) and chloride (Cl - ). We show that active Cld enzymes can be targeted to either the cytosol or mitochondria of human cells, and that coexpressing a chlorite transporter results in molecular oxygen production inside cells in response to externally added chlorite. This first-generation system allows fine temporal and spatial control of oxygen production, with immediate research applications. In the future, we anticipate that technologies based on SNORCL will have additional widespread applications in research, biotechnology, and medicine. |
