Studying Plant MIF/D-DT-Like Genes and Proteins (MDLs).

Human macrophage migration inhibitory factor (MIF) is an inflammatory cytokine with chemokine-like characteristics and an upstream regulator of host innate immunity. It is a critical mediator of a variety of human diseases, such as acute and chronic inflammatory diseases, autoimmunity, atherosclerosis, and cancer. MIF is an atypical chemokine that not only signals through its cognate receptor CD74, but also interacts with the classical chemokine receptors CXCR2 and CXCR4. MIF and its homolog D-dopachrome tautomerase (D-DT)/MIF-2 are structurally unique proteins that are conserved across kingdoms and that share a remarkable homology with bacterial tautomerases/isomerases, albeit the relevance of the tautomerase activity in mammalian systems has remained unclear. Intriguingly, in silico analysis also predicts MIF orthologs in plants such as in the model plant Arabidopsis thaliana. There are three predicted MIF orthologs in A. thaliana, which have been termed A. thaliana MIF/D-DT-like proteins (AtMDLs). Anticipating that there will be a future research interest in studying AtMDLs or other plant MDLs, here we describe methods how to clone, recombinantly express and purify AtMDL proteins, taking into account codon usage differences between plant and mammalian cell systems.