Expression, purification, and encephalitogenicity of recombinant human myelin oligodendrocyte glycoprotein.
Myelin oligodendrocyte glycoprotein (MOG), a putative autoantigen in multiple sclerosis (MS), is a quantitatively minor component of the CNS. In view of the difficulties associated with the purification of MOG from brain tissues, the extracellular domain of human MOG corresponding to the N-terminal 121 amino acids was expressed in Escherichia coli as a glutathione sulfotransferase fusion protein. The expressed protein was localized to inclusion bodies, and varying the growth parameters resulted in the solubilization of small amounts of GST-MOG that could be affinity purified on glutathione agarose columns. The fusion protein found in the inclusion bodies could be solubilized with urea. The solubilized fusion protein was cleaved with thrombin, and the extracellular domain was purified by CM Sephadex 50 chromatography to homogeneity. Injection of recombinant human MOG into different strains of mice resulted in the induction of an MS-like disease, characterized by severe neurological impairment and extensive CNS demyelinated lesions. Recombinant MOG produced in E. coli should prove to be useful as a highly purified biological reagent for immunological, pathological, functional, and structural studies.
Bettadapura
J
J
Neuroimmunology Laboratory, La Trobe University, Bundoora, Victoria, Australia.
Menon
K K
KK
Moritz
S
S
Liu
J
J
Bernard
C C
CC
D000818:Animals; D004660:Encephalitis / Q000139:chemically induced*; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D051379:Mice; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D063308:Myelin-Oligodendrocyte Glycoprotein; D011994:Recombinant Proteins; D012995:Solubility; D014508:Urea / Q000494:pharmacology
C571326:MOG protein, human; C571327:Mog protein, mouse; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein; D011994:Recombinant Proteins; D014508:Urea; D005982:Glutathione Transferase