Two-dimensional high-resolution electrophoresis of elastin-derived peptides

Degradation of human aortic elastin in vivo yields a restricted number of differentially sized and charged peptides. Elastin-derived peptides (EDP) can be analyzed by two-dimensional electrophoresis after their extraction from human abdominal aortic tissue by 0.2 M sodium chloride. The peptides were separated according to charge by acetic acid-urea-PAGE and then according to molecular mass by SDS-PAGE. The identity of these peptides as EDP was continued by immunoprecipitation and Western blots. The two-dimensional electrophoretic system can resolve desmosine-like cross-linked EDP of the similar molecular configuration but differing in the number of positively charged amino acid residues. The new separation technique of EDP has the capacity to identify defects in desmosine-like cross-links and may be useful in characterizing abberations in elastin structures.