Identification of macrophage migration inhibitory factor isoforms in bovine brain.
In the course of the study of the primary structures and molecular mechanisms of action of immunologically active compounds of the nervous system we have isolated from the soluble fraction of total bovine brain two heat-stable proteins. The purification procedure was mainly based on DEAE-Servacel ion-exchange chromatography and reversed-phase HPLC. The proteins were identified by the N-terminal Edman microsequence analysis and database searching as macrophage migration inhibitory factor (MIF). The N-terminal sequences for MIF1 and MIF2 were found to be identical. According to mass spectral analysis, the molecular masses for MIF1 and MIF2 were determined respectively as 12,369.21 and 12,299.7 Da. In addition, we have also isolated a third peptide having the same N-terminal sequence and Mr 9,496.2 that seems to be a proteolytic fragment of MIF. Using p-hydroxyphenylpyruvate as a substrate, we have not revealed tautomerase activity of either MIF1 or MIF2. As both the immunologic and enzymatic activities were reported to be expressed by the oligomeric structure of MIF, we suggest that the present study may give additional information on MIF in terms of structural properties of this protein. A comparatively simple purification procedure is presented that may be widely used for simultaneous isolation in one run of MIF isoforms.
Gurvits
B Y
BY
Institute of Biochemistry, Russian Academy of Sciences, Moscow. inbio@glas.apc.org
Tretyakov
O Y
OY
Klishina
N V
NV
Stoeva
S
S
Voelter
W
W
Galoyan
A A
AA
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D008263:Macrophage Migration-Inhibitory Factors / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification
D008263:Macrophage Migration-Inhibitory Factors; D010446:Peptide Fragments; D020033:Protein Isoforms