Affinity Purification of Macrophage Migration Inhibitory Factor/Glycosylation Inhibiting Factor (MIF/GIF) from Bovine Brain by Using a Peptide Ligand Derived from a Novel Serpin

Abstract

We purified macrophage migration inhibitory factor/glycosylation inhibiting factor (MIF/GIF) from bovine brain by using an affinity column with the C-terminal region peptide of a novel serpin as a ligand. The affinity purified preparation showing a single band on SDS-PAGE contained four peptides on RP-HPLC, which were converged into two peptides time-dependently. Sequence analysis and Western blotting revealed that one was identical to bovine MIF/GIF and the other was an N-terminally modified form of MIF/GIF. These results indicated that there exist at least two forms of MIF/GIF in the bovine brain and that they have an affinity for the C-terminal portion of the serpin.