FIGURE 5.

Glycation of ATP ghosts. The ghosts were prepared by lysis of cells in the presence or absence of ATP and resealed as described under “Methods.” These ghosts were glycated with ribose at 37 °C for up to 60 min. A, membrane proteins were separated on 5% SDS-PAGE followed by immunoblotting with anti-pentosidine antibody. B, membrane deformability of these ghosts was measured by the ektacytometry. C, flippase inhibitor, PDA or N-ethylmaleimide (NEM), was incorporated into ATP ghosts. Pentosidine formation of spectrin molecules was analyzed by SDS-PAGE. Gels were stained with CBB, blotted onto PVDF, and probed with anti-pentosidine antibody.