Fig. 3.

Analysis of major MBP cleavage products. Reverse-phase HPLC-MS analysis of major MBP cleavage products. Column eluate fractions, corresponding to dominant chromatography peaks (A), were collected, freeze dried, redissolved, and applied to SELDI H4 chip and tricine-SDS/PAGE (B). Gel was stained by Sypro Orange. Peptides, unambiguously identified by SELDI and clearly seen in corresponding gel lanes, are indicated. (C) Schematic description of the preferential antibody cleavage sites in the MBP sequence. Sequence fragments identical to the immunodominant MBP-derived peptides (12-31, 82-98, 110-128, and 144-169) are shown in yellow rectangles, and the encephalitogenic peptide region (86-98) is marked by the red box. (D) ³²P-MBP degradation by autoantibodies. Autoradiography of ³²P-phosphorylated MBP hydrolysis by proteolytic mouse (EAE SJL) and human (MS) IgG. Line M, molecular mass markers (range 2.5-16.9 kDa, Amersham Pharmacia).