Presentation and detection of azide functionality in bacterial cell surface proteins.
An improved protocol for copper-catalyzed triazole formation on the bacterial cell surface is described. Addition of highly pure CuBr to cells treated with azidohomoalanine (2) leads to ca. 10-fold more extensive cell surface labeling than previously observed. This highly active catalyst allows detection of the methionine analogues azidoalanine (1), azidonorvaline (3), and azidonorleucine (4) in cell surface proteins. Azidoalanine was previously believed to be silent with regard to the cellular protein synthesis machinery.
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
A James
AJ
Link
Mandy K S
MK
Vink
David A
DA
Tirrell
D000409:Alanine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000378:metabolism; D001386:Azides / Q000032:analysis* / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000032:analysis / Q000378:metabolism; D005434:Flow Cytometry; D008715:Methionine / Q000378:metabolism; D009646:Norleucine / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014230:Triazoles / Q000032:analysis* / Q000378:metabolism; D014633:Valine / Q000031:analogs & derivatives* / Q000378:metabolism
D001386:Azides; D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D014230:Triazoles; D003300:Copper; D009646:Norleucine; C039309:azidoalanine; C005313:norvaline; D008715:Methionine; D014633:Valine; D000409:Alanine