Rapid method based on reversed-phase high-performance liquid chromatography for purification of human myelin basic protein and its thrombic and endoproteinase Lys-C peptides.
Reversed-phase high-performance liquid chromatography was applied to isolate myelin basic protein from human brain, followed by separation of proteolytic peptides thereof on the same chromatographic system. Brain tissue was delipidated under conditions that keep copurifying proteases inactive. The crude brain protein fraction was applied directly to a C4 column. The homogeneous protein obtained in this way was digested with thrombin and endoproteinase Lys-C in order to produce short defined myelin basic protein peptides. The purified peptides were used to determine the antigen fine specificity of myelin basic protein recognizing T lymphocyte lines isolated from multiple sclerosis patients.
Max-Planck-Institut für Psychiatrie, Martinsried, F.R.G.
G
G
Giegerich
M
M
Pette
K
K
Fujita
H
H
Wekerle
J T
JT
Epplen
A
A
Hinkkanen
D001923:Brain Chemistry; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D010450:Endopeptidases; D006801:Humans; D008213:Lymphocyte Activation; D008666:Metalloendopeptidases; D009103:Multiple Sclerosis / Q000276:immunology; D004676:Myelin Basic Protein / Q000276:immunology / Q000302:isolation & purification*; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000276:immunology / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000276:immunology; D013917:Thrombin
D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D010450:Endopeptidases; D013917:Thrombin; D008666:Metalloendopeptidases; C039830:peptidyl-Lys metalloendopeptidase