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A high-throughput fluorescence assay for amidases is demonstrated. It illustrates a new principle in enzyme assays, whereby changes in the pM value during the reaction are recorded (pM=−lg[M], M=free metal ions). An orange fluorescent quinacridone-based ligand 1 is the pM sensor (shown chelated to a reporter metal ion, M2+=Cu2+ or Ni2+) that is used to monitor the release of metal-chelating amino acids by the enzymes acylase I and aminopeptidase.
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Klein et al. (2001) studied this question.
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