TY - JOUR
T1 - Steady-state fluorescence emission from the fluorescent probe, 5-iodoacetamidofluorescein, bound to hemoglobin
AU - Hirsch, Rhoda Elison
AU - Zukin, R. Suzanne
AU - Nagel, Ronald L.
N1 - Funding Information:
This project was supported in part by a New York Heart Association J. Fred Weintz Investigatorship (to R.E.H.), Cottrell College Science Grant (Research Corp.) No.1281 (to R.E.H.), and NIH Grant AL21016 (to R.E.H. & R.L.N.).
PY - 1986/7/16
Y1 - 1986/7/16
N2 - In the past, fluorescence emission from an extrinsic fluorophore bound to heme-proteins would only be studied with the removal of the heme since fluorescence from the flurophore could not be detected using right-angle optics. Using front-face fluorometry, a significant steady state emission signal originating from the probe bound to hemoglobin is detected. This is the first report of the detection of extrinsic fluorescence of a probe bound to a heme-protein. We also demonstrate that the extrinsic probe, 5-iodoacetamidofluorescein, is covalently bound to hemoglobin, specifically at β93 Cysteine. Ligand binding results in a change in the fluorophore fluorescence intensity as predicted by hemoglobin crystallographic studies. Efficiency of energy transfer measurements are made.
AB - In the past, fluorescence emission from an extrinsic fluorophore bound to heme-proteins would only be studied with the removal of the heme since fluorescence from the flurophore could not be detected using right-angle optics. Using front-face fluorometry, a significant steady state emission signal originating from the probe bound to hemoglobin is detected. This is the first report of the detection of extrinsic fluorescence of a probe bound to a heme-protein. We also demonstrate that the extrinsic probe, 5-iodoacetamidofluorescein, is covalently bound to hemoglobin, specifically at β93 Cysteine. Ligand binding results in a change in the fluorophore fluorescence intensity as predicted by hemoglobin crystallographic studies. Efficiency of energy transfer measurements are made.
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U2 - 10.1016/0006-291X(86)90307-4
DO - 10.1016/0006-291X(86)90307-4
M3 - Article
C2 - 3755598
AN - SCOPUS:0022455764
SN - 0006-291X
VL - 138
SP - 489
EP - 495
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -