Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles.
The E. coli aminoacyl transferase (AaT) can be used to transfer a variety of unnatural amino acids, including those with azide or alkyne groups, to the α-amine of a protein with an N-terminal Lys or Arg. Subsequent functionalization through either copper-catalyzed or strain-promoted click reactions can be used to label the protein with fluorophores or biotin. This method can be used to directly detect AaT substrates or in a two-step protocol to detect substrates of the mammalian ATE1 transferase.
Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA, 19104-6323, USA.
Anne M
AM
Wagner
John B
JB
Warner
Haviva E
HE
Garrett
Christopher R
CR
Walters
E James
EJ
Petersson
D019881:Aminoacyltransferases / Q000378:metabolism; D001126:Arginine-tRNA Ligase / Q000378:metabolism; D002384:Catalysis; D057930:Click Chemistry / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins; D013194:Staining and Labeling; D014166:Transferases / Q000378:metabolism*
D011506:Proteins; D011994:Recombinant Proteins; D014166:Transferases; D019881:Aminoacyltransferases; D001126:Arginine-tRNA Ligase