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

Authors

• Andrew Parkins, University of the PacificFollow
• Georgios Pantouris, University of the PacificFollow

Document Type

Article

Publication Title

STAR Protocols

Department

Chemistry

ISSN

26661667

Volume

4

Issue

3

DOI

10.1016/j.xpro.2023.102375

First Page

1

Last Page

12

Publication Date

9-15-2023

Abstract

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

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Parkins, Andrew and Pantouris, Georgios, "Protocol for purification and enzymatic characterization of members of the human macrophage migration inhibitory factor superfamily" (2023). Pacific Faculty Work. 133.
https://scholarlycommons.pacific.edu/all-faculty/133