Site-specific incorporation of fluorotyrosines into proteins in Escherichia coli by photochemical disguise.
Fluorinated analogues of tyrosine can be used to manipulate the electronic environments of protein active sites. The ability to selectively mutate tyrosine residues to fluorotyrosines is limited, however, and can currently only be achieved through the total synthesis of proteins. As a general solution to this problem, we genetically encoded the unnatural amino acids o-nitrobenzyl-2-fluorotyrosine, -3-fluorotyrosine, and -2,6-difluorotyrosine in Escherichia coli. These amino acids are disguised from recognition by the endogenous protein biosynthetic machinery, effectively preventing global incorporation of fluorotyrosine into proteins.
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Bryan J
BJ
Wilkins
Samuel
S
Marionni
Douglas D
DD
Young
Jia
J
Liu
Yan
Y
Wang
Martino L
ML
Di Salvo
Alexander
A
Deiters
T Ashton
TA
Cropp
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015394:Molecular Structure; D015202:Protein Engineering / Q000379:methods; D017433:Protein Structure, Secondary; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
D029968:Escherichia coli Proteins; C001245:3-fluorotyrosine; D014443:Tyrosine; C060489:2-fluorotyrosine