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Figure 1 | BMC Biochemistry

Figure 1

From: Role of redox environment on the oligomerization of higher molecular weight adiponectin

Figure 1

Re-oligomerization of adiponectin following dialysis into defined concentrations of DTT. (A) Native PAGE of adiponectin oligomers after collapse to trimers and subsequent re-oligomerization as concentrations of DTT were equilibrated from 5 mM to 0.008, 0.04, 0.2, 1, or 5 mM by dialysis over an 18-hr period. (B) Non-reducing denaturing SDS-PAGE analysis to identify the oxidation state of adiponectin as either in the disulfide-bonded dimer state or reduced monomer state in each of the different conditions. Oligomerization and oxidation states were examined at 6 and 18 hrs. The redox state of adiponectin was fixed by adding NEM to the sample at the end of reassembly reactions. 3 mer: trimer; 6 mer: hexamer; 9 mer: nonamer; 12 mer: dodecamer; 18 mer: octadecamer. The number of monomers in trimers, hexamers, and octadecamers in native gels were independently verified using native mass spectrometry and gel filtration chromatography (data not shown). The number of monomers in nonamers and dodecamers were determined by extrapolation of migration distances of known oligomers in native gels as described in Methods.

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