Distribution of fluorescence decay times for 1-anilinonaphthalene-8-sulfonate in human oxyhemoglobin A1b solution

We have studied complex formation between molecules of the fluorescent probe 1-anilinonaphthalene-8-sulfonate (1,8-ANS) and the major form (A1) and a minor form (Ab) of hemoglobin. The contribution of the longlived component f3 to the kinetic curves for fluorescence decay in HbA1b solutions is 0.021–0.036, which indicates a dramatic decrease (compared with HbA1) in the accessibility of the central cavity of HbA1b for binding 1,8-ANS. Disappearance of the long-lived component f3 in the fluorescence decay kinetics of 1,8-ANS in HbA1b solutions in the presence of inositol hexaphosphate (IHP) suggests that the regulatory region of HbA1b is completely inaccessible for interaction both with the negatively charged molecules of the probe and with natural regulators of the transport function for this form of the heme protein.