Characterization of a recombinant murine 18.5-kDa myelin basic protein.
A recombinant hexahistidine-tagged 18.5-kDa isoform of murine myelin basic protein has been characterized biochemically and immunogenically, by mass spectrometry, by circular dichroism under various conditions (in aqueous solution, with monosialoganglioside G(M1), and in 89% 2-propanol), and by transmission electron microscopy. The preparations of this protein indicated a high degree of purity and homogeneity, with no significant posttranslational modifications. Circular dichroic spectra showed that this preparation had the same degree of secondary structure as the natural bovine 18.5-kDa isoform of myelin basic protein. Incubation of the recombinant protein with lipid monolayers containing a nickel-chelating lipid resulted in the formation of fibrous assemblies that formed paracrystals of spacings 4.8 nm between fibers and 3-4 nm along them.
Bates
I R
IR
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Matharu
P
P
Ishiyama
N
N
Rochon
D
D
Wood
D D
DD
Polverini
E
E
Moscarello
M A
MA
Viner
N J
NJ
Harauz
G
G
D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli; D005732:Gangliosides / Q000378:metabolism; D050356:Lipid Metabolism; D008055:Lipids; D013058:Mass Spectrometry; D051379:Mice; D008854:Microscopy, Electron; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000648:ultrastructure; D009532:Nickel / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000648:ultrastructure; D013601:T-Lymphocytes / Q000276:immunology
C109930:2-(bis-carboxymethylamino)-6-(2-(1,3-di-O-oleylglyceroxy)acetylamino)hexanoic acid nickel(II); D000939:Epitopes; D005732:Gangliosides; D008055:Lipids; D004676:Myelin Basic Protein; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C025447:sialogangliosides; D009532:Nickel