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Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine implicated in the pathogenesis of multiple acute and chronic inflammatory conditions, harbours diverse catalytic activity, notably the ability to tautomerize 2-carboxy-2,3-dihydroxyindole-5,6-quinone (L-dopachrome) to its dihydroxyindole derivative. While an endogenous substrate for this tautomerase activity is yet to be fully characterized, it is possible MIF enzymatic activity is important for numerous biological functions. As a result, assessing the tautomerase activity of MIF may serve as a useful screening tool for identifying small-molecule MIF inhibitors. This chapter details the use of a simple, cell-free, spectrophotometric assay for investigating the inhibitory potential of small-molecule compounds against the keto-enol tautomerase activity of MIF.

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Assessing the Tautomerase Activity of MIF Using L-Dopachrome

  • 实验方案

  • 发表日期：

    2026年

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  • Katie Treasure,
  • Amanda Genders,
  • James Harris

摘要

Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine implicated in the pathogenesis of multiple acute and chronic inflammatory conditions, harbours diverse catalytic activity, notably the ability to tautomerize 2-carboxy-2,3-dihydroxyindole-5,6-quinone (L-dopachrome) to its dihydroxyindole derivative. While an endogenous substrate for this tautomerase activity is yet to be fully characterized, it is possible MIF enzymatic activity is important for numerous biological functions. As a result, assessing the tautomerase activity of MIF may serve as a useful screening tool for identifying small-molecule MIF inhibitors. This chapter details the use of a simple, cell-free, spectrophotometric assay for investigating the inhibitory potential of small-molecule compounds against the keto-enol tautomerase activity of MIF.