Metal Substitution and Fluorescence Modifications in Ascorbate-Alkaline Phosphatase Interaction
Alkaline phosphatase (EC 220.127.116.11), a zinc metallo-enzyme, is irreversibly inhibited by ascorbate, probably through a modification of active site environment (Martorana et al., 1986; Miggiano et al., 1984). A metal substituted form was prepared from bovine kidney alkaline phosphatase purchased from Calbiochem (La Jolla, CA, U.S.A.) and further purified as described (Martorana et al., 1986). Cobalt can replace zinc yielding a partially active phosphohydrolase, which lacks phosphotransferase activity (Ensinger et al., 1978) and presents a relevant visible absorption (Simpson and Vallee, 1968).
KeywordsZinc Chloride Charge Transfer Mechanism Free Tryptophan Difference Absorption Spectrum Cobalt Sulfate
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