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. Author manuscript; available in PMC: 2010 Dec 29.
Published in final edited form as: Biochemistry. 2009 Dec 29;48(51):12345–12357. doi: 10.1021/bi9015555

Figure 2. Effect of oxidizing agent hydrogen peroxide on (A) re-oligomerization of octadecameric adiponectin and (B) oxidation of cysteines near N-terminus to form disulfide bonds.

Figure 2

Purified adiponectin octadecamers were first reduced by DTT followed by lowering the pH to 5 as described in Experimental Procedures to produce trimers. The samples were subsequently dialyzed against PBS at pH 7.6 to remove DTT in the absence or presence of 5 mM hydrogen peroxide over a period for 3 hrs as indicated. Samples were removed at different time points during the dialysis and fractionated in native gels to monitor oligomerization states and in non-reducing denaturing gels to distinguish the oxidized (dimer) and reduced (monomer) forms of adiponectin. Oxidation states were fixed by NEM treatment.