Fig. 4.

Analysis of EGS-cross-linked stomatin-complexes in DRMs. Stripped ghosts were treated with cold Triton X-100 and subjected to flotation. DRMs were isolated and cross-linked with 16 μM EGS. After quenching and solubilisation with SDS, the mixture was adjusted to 1% Triton X-100 and stomatin-complexes were immunopurified. Elution fractions were analysed by (A) 10% or (B) 7% SDS-PAGE/silver staining. Stained bands were excised, as indicated, and analysed by MS. (C) Semi-quantitative results (AvTIC units) show the size-distribution of stomatin, band 3, and GLUT1. Stomatin peaks are clearly seen at the size of oligomers ( ≥ 500 kDa), the homo-dimer (65 kDa), and monomer (30 kDa). The 20 kDa stomatin may represent an intramolecularly cross-linked species. Low-molecular band 3 may represent fragments or contamination. M, marker; 1–5, respective elution fractions.