Biological activity of recombinant human myelin basic protein.
Using an inducible expression vector in Escherichia coli, we have expressed, purified, and biologically characterized recombinant human myelin basic protein (r-MBP). The recombinant protein binds in cation-exchange chromatography with similar affinity to purified, human MBP, and elutes as a single, 18.5-kDa species as judged by SDS-PAGE. The recombinant protein exhibits similar conformation to native MBP, as demonstrated by ELISA reactivity with a panel of six monoclonal antibodies directed against at least three different epitopes on human MBP. Additionally, recombinant MBP can trigger the proliferation of human T cell clones recognizing MBP, can induce experimental autoimmune encephalomyelitis (EAE) in the SJL mouse, and is capable of suppressing EAE following tolerization via oral administration. Most important, by using a novel method of purification, the recombinant protein preparation contains no detectable proteolytically derived fragments of MBP, a significant advantage over MBP purified from protease-rich central nervous system tissue. Purified recombinant MBP produced in E. coli will be useful as a biological reagent where highly purified protein devoid of degradation products is needed. Relevance to the study of antigen processing, interactions between MHC and the TCR, as well as for the investigation of antigen-driven immune tolerance are discussed.
Oettinger
H F
HF
Department of Medicine, Brigham & Women's Hospital, Boston, MA.
al-Sabbagh
A
A
Jingwu
Z
Z
LaSalle
J M
JM
Weiner
H L
HL
Hafler
D A
DA
D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000209:etiology / Q000517:prevention & control; D005260:Female; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D004676:Myelin Basic Protein; D011994:Recombinant Proteins