Soil nitrogen is the most important element affecting the growth and yield of crops and is also a key component of soil organic matter. Despite great efforts that have been made, the nature of about 50% of nitrogen in...Soil nitrogen is the most important element affecting the growth and yield of crops and is also a key component of soil organic matter. Despite great efforts that have been made, the nature of about 50% of nitrogen in soils and humic acids still remains obscure. Usually called unknown nitrogen, such nitrogen might be divided into two categories, the hydrolyzable nitrogen (HUN) and the nonhydrolyzable nitrogen.展开更多
Studies on the mechanism of protein phosphorylation and therapeutic interventions of its related molecular processes are limited by the difficulty in the production of purpose-built phosphoproteins harboring site-spec...Studies on the mechanism of protein phosphorylation and therapeutic interventions of its related molecular processes are limited by the difficulty in the production of purpose-built phosphoproteins harboring site-specific phosphorylated amino acids or their nonhydrolyzable analogs.Here we address this limitation by customizing the cell-free protein synthesis(CFPS)machinery via chassis strain selection and orthogonal translation system(OTS)reconfiguration screening.The suited chassis strains and reconfigured OTS combinations with high orthogonality were consequently picked out for individualized phosphoprotein synthesis.Specifically,we synthesized the sfGFP protein and MEK1 protein with site-specific phosphoserine(O-pSer)or its nonhydrolyzable analog,2-amino-4-phosphonobutyric acid(C-pSer).This study successfully realized building cell-free systems for site-specific incorporation of phosphonate mimics into the target protein.Our work lays the foundation for developing a highly expansible CFPS platform and the streamlined production of user-defined phosphoproteins,which can facilitate research on the physiological mechanism and potential interference tools toward protein phosphorylation.展开更多
基金Project supported by the National Natural Science Foundation of China
文摘Soil nitrogen is the most important element affecting the growth and yield of crops and is also a key component of soil organic matter. Despite great efforts that have been made, the nature of about 50% of nitrogen in soils and humic acids still remains obscure. Usually called unknown nitrogen, such nitrogen might be divided into two categories, the hydrolyzable nitrogen (HUN) and the nonhydrolyzable nitrogen.
基金supported by the National Natural Science Foundation of China (21878173,22177059)National Key R&D Program of China (2018YFA0901700,2021YFC2103900)a grant from the Institute Guo Qiang,Tsinghua University (2021GQG1016).
文摘Studies on the mechanism of protein phosphorylation and therapeutic interventions of its related molecular processes are limited by the difficulty in the production of purpose-built phosphoproteins harboring site-specific phosphorylated amino acids or their nonhydrolyzable analogs.Here we address this limitation by customizing the cell-free protein synthesis(CFPS)machinery via chassis strain selection and orthogonal translation system(OTS)reconfiguration screening.The suited chassis strains and reconfigured OTS combinations with high orthogonality were consequently picked out for individualized phosphoprotein synthesis.Specifically,we synthesized the sfGFP protein and MEK1 protein with site-specific phosphoserine(O-pSer)or its nonhydrolyzable analog,2-amino-4-phosphonobutyric acid(C-pSer).This study successfully realized building cell-free systems for site-specific incorporation of phosphonate mimics into the target protein.Our work lays the foundation for developing a highly expansible CFPS platform and the streamlined production of user-defined phosphoproteins,which can facilitate research on the physiological mechanism and potential interference tools toward protein phosphorylation.
