Photolabeling of human serum albumin by 4-azido-2-([14C]-methylamino) trifluorobenzonitrile. A high-efficiency, long wavelength photolabel.
N-alkyl derivatives of perfluoroarylazides are excellent candidates for photolabeling of proteins since they have absorption spectra in the 340-350 nm range permitting photolabel absorption without direct protein photolysis. The [14C]-N-methylamino derivative of 4-azido-tetrafluorobenzonitrile has been used to demonstrate that 80% of the photo-induced nitrene transient becomes covalently attached to HSA during photolysis. Multiwavelength detection of the photoprobe-protein separation by size exclusion chromatography is shown to be an effective tool for assessing the conjugation of the photoprobe to the protein.
Pandurangi
R S
RS
Department of Chemistry, University of Missouri, Columbia, USA.
Kuntz
R R
RR
Volkert
W A
WA
D000345:Affinity Labels / Q000138:chemical synthesis / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002250:Carbon Radioisotopes; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D009570:Nitriles / Q000737:chemistry; D010782:Photolysis; D012709:Serum Albumin / Q000737:chemistry*
C094048:4-azido-2-(methylamino)trifluorobenzonitrile; D000345:Affinity Labels; D001386:Azides; D002250:Carbon Radioisotopes; D009570:Nitriles; D012709:Serum Albumin