Quenching of alkaline phosphatase phosphorescence by O2 and NO. Evidence for inflexible regions of protein structure.
The rate constant for quenching the phosphorescence of alkaline phosphatase by molecular oxygen was measured as a function of temperature. The results disagree with previous determinations and, contrary to fluorescence quenching, show that diffusion of O2 to this region of the macromolecule is a highly hindered process. When nitric oxide is introduced as a quencher, similarly small rate constants were found. While the activation energy for this process is identical for both quenchers, it is much smaller than for structural fluctuations at the chromophore site as manifested by the intrinsic triplet-state lifetime. These findings are analyzed in terms of a mechanism that takes into account static quenching at large distances and does not require penetration of the quencher all the way to the chromophore.
Strambini
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D000469:Alkaline Phosphatase / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D049449:Luminescence; D009569:Nitric Oxide / Q000494:pharmacology*; D010100:Oxygen / Q000494:pharmacology*; D011487:Protein Conformation; D013816:Thermodynamics
D009569:Nitric Oxide; D000469:Alkaline Phosphatase; D010100:Oxygen