A fluorometric assay for pyridoxal kinase applicable to crude cell extracts.
We have developed an assay for pyridoxal kinase which takes advantage of the intense fluorescence yield of the oxime of pyridoxal or pyridoxal-5'-phosphate and the substantial difference in the rates of formation of these two oximes. Evidence is presented which demonstrates that the assay is linear with respect to time and amount of protein and is applicable to the activity in crude cell extracts as well as purified enzyme.
Sussmane
S
S
Department of Biochemistry, University of Tennessee, Knoxville 37996, USA.
Koontz
J
J
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D019811:Hydroxylamine; D006898:Hydroxylamines; D007202:Indicators and Reagents; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D011730:Pyridoxal / Q000031:analogs & derivatives / Q000032:analysis; D011731:Pyridoxal Kinase / Q000032:analysis* / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000032:analysis; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods; D014407:Tumor Cells, Cultured
D006898:Hydroxylamines; D007202:Indicators and Reagents; D019811:Hydroxylamine; D011730:Pyridoxal; D011732:Pyridoxal Phosphate; C028073:pyridoxaloxime; D011731:Pyridoxal Kinase