Characterization of the H- and L-subunit ratios of ferritins by sodium dodecyl sulfate-capillary gel electrophoresis.
Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) was used to characterize the H- and L-subunit ratios of several mammalian ferritins and one bacterioferritin. Traditionally, SDS-PAGE has been used to characterize the H- and L-subunit ratios in ferritin; however, this technique is relatively slow and requires staining, destaining, and scanning before the data can be processed. In addition, the H- and L-subunits of ferritin are fairly close in molecular weight (approximately 21,000 and approximately 20,000, respectively) and are often difficult to resolve in SDS-PAGE slab gels. In contrast, SDS-CGE requires no staining or destaining procedures and the peak quantitation is superior to SDS-PAGE. SDS-CGE is effective in quickly resolving the H- and L-subunits of ferritins from horse spleen, human liver, recombinant human H and L homopolymers, and mixtures of the two- and the single-subunit of a bacterioferritin from Escherichia coli. The technique has also proven useful in assaying the quality of the protein sample from both commercial and recombinant sources. Significant amounts of low-molecular-weight degradation products were detected in all commercial sources of horse spleen ferritin. Most commercial horse spleen ferritins lacked intact H-subunits under denaturing conditions.
Grady
John K
JK
Department of Chemistry, University of New Hampshire, Rudman Hall, Durham, New Hampshire 03824, USA.
Zang
Jia
J
Laue
Thomas M
TM
Arosio
Paolo
P
Chasteen
N Dennis
ND
D000818:Animals; D001426:Bacterial Proteins; D003573:Cytochrome b Group / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli; D005293:Ferritins / Q000032:analysis*; D006736:Horses; D006801:Humans; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000032:analysis
D001426:Bacterial Proteins; D003573:Cytochrome b Group; D021122:Protein Subunits; D011994:Recombinant Proteins; D005293:Ferritins; C018032:bacterioferritin