Anomalous Electrophoretic Behavior of a Cyanogen Bromide Peptide from Type III Collagen

Abstract

The relationship between relative mobility on sodium dodecyl sulfate-polyacrylamide gels and the logarithm of molecular weight was linear for peptides derived by CNBr cleavage from purified collagen chains. One striking exception was found: when the CNBr peptide alpha 1(III)CB-8 was subjected to electrophoresis on a polyacrylamide slab gel calibrated with other peptides derived from the type III collagen chain, its relative mobility corresponded to a molecular weight of approximately 8,000. On a gel calibrated with globular proteins, its apparent molecular weight was 12,000. Its actual molecular weight, as determined by gel filtration and by amino acid analysis, is approximately 12,000. The dodecyl sulfate-protein binding ratio for this peptide did not differ from that of other collagenous peptides or globular proteins. Free electrophoretic mobilities and retardation coefficients were determined for selected collagenous peptides and globular proteins by electrophoresis on tube gels of different concentrations of acrylamide. By these indices, alpha 1(III)CB-8 resembled a globular protein rather than a collagenous peptide. Presumably, some difference in primary structure of this peptide results in conformational changes of its complex with dodecyl sulfate, such that the peptide-dodecyl sulfate complex behaves more like a globular protein than a collagenous peptide on polyacrylamide gels.