L-4-tetrazine-phenylalanine Synthesis And Its Applications In The Bioorthogonal Reactions

The study of biomolecules in their native environment is a dream of the scientists, but it is a challenging task because of the vast complexity of cellular systems. Technologies developed in the last few years for the selective modification of biological species in living systems have yielded new insights into cellular processes. Key to these new techniques are bioorthogonal chemical reactions, whose components must react rapidly and selectively with each other under physiological conditions in the presence of the plethora of functionality. The Cu-catalyzed azide-alkynel 1,3-dipolar cycloaddition reaction is a typical bioorthogonal chemistry. However, the need for a copper catalyst, which is known to be toxic to cells, precludes its potential applications in live cells.Then several strain-promoted azide-alkynel cycloaddition have been developed to label biomolecules in vivo. Base on Inverse-Electron-Demand Diels-Alder Reactivity of tetrazine and norbornene or trans-cyclooctene reactivity is another selectivity. The reactions tolerate a broad range of functionality and proceed in high yield in organic solvents,water,cell media,or cell lysate. So, if proteins, glycans, lipids, nucleic acids fashioned with tetrazine, then biotin, fluorophores, antibody can probe them,tacking and study these biomolecules’localization, transportation, metabolism etc. So, we used L-4-CN-phenylalanine synthesis L-4-1,2,4,5-tetrazine-phenylalanine.In PBS, norbornene can fast get reaction with L-4-1,2,4,5-tetrazine-phenylalanine. This can demonstrate through genetic encoding,replace phenylanine or tyrosine by L-4-1,2,4,5-tetrazine-phenylalanine, then the probe carried nobornene can image proteins by bioorthogonal reaction.