Characterization of the H- and L-Subunit Ratios of Ferritins by Sodium Dodecyl Sulfate–Capillary Gel Electrophoresis

✍ Scribed by John K. Grady; Jia Zang; Thomas M. Laue; Paolo Arosio; N.Dennis Chasteen

• Publisher: Elsevier Science
• Year: 2002
• Tongue: English
• Weight: 78 KB
• Volume: 302
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2001.5561

✦ Synopsis

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 (ϳ21,000 and ϳ20,000, 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.