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  • Title: Fluorescent labeling of (Na+ + K+)-ATPase by 5-iodoacetamidofluorescein.
    Author: Kapakos JG, Steinberg M.
    Journal: Biochim Biophys Acta; 1982 Dec 22; 693(2):493-6. PubMed ID: 6297560.
    Abstract:
    5-Iodoacetamidofluorescein (5-IAF) covalently labels dog kidney (Na+ + K+)-ATPase with approximately 2 moles incorporated per mole of enzyme. ATPase and K+-phosphatase activities are fully retained after reaction, and the kinetic parameters for Na+, K+, Mg2+, ATP and p-nitrophenyl phosphate are likewise not significantly affected. The fluorescence of the bound 5-IAF is increased by ATP, Na+, and Mg2+, and decreased by K+. These fluorescence changes likely reflect ligand-induced stabilization of the E1 or E2 states of the enzyme.
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