Protocol for purification and enzymatic characterization of members of the human macrophage migration inhibitory factor superfamily

ABSTRACT: Summary Macrophage migration inhibitory factor (MIF) and D-dopachrome tautomerase (D-DT or MIF-2) are two proteins serving a key role in the pathogenesis of multiple disorders, including cancer.1 Here, we present a protocol for the purification and enzymatic characterization of MIF and D-DT using keto-enol tautomerase activity. This approach measures enzymatic activity through the formation of an enol-borate complex. We describe steps for expressing and purifying proteins, preparing the 96-well microplate, and assay implementation including monitoring of keto-enol tautomerase activity. For complete details on the use and execution of this protocol, please refer to Parkins et al.2,3 Graphical abstract Highlights • Systematic protocol for the purification and enzymatic characterization of M

SUBMITTER: Parkins A 

PROVIDER: S-EPMC10319315 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature