title: Zinc enhances adiponectin oligomerization to octadecamers but decreases the rate of disulfide bond formation.
abstract: Adiponectin, a hormone secreted from adipocytes, has been shown to protect against development of insulin resistance, ischemia-reperfusion injury, and inflammation. Adiponectin assembles into multiple oligomeric isoforms: trimers, hexamers and several higher molecular weight (HMW) species. Of these, the HMW species are selectively decreased during the onset of type 2 diabetes. Despite the critical role of HMW adiponectin in insulin responsiveness, its assembly process is poorly understood. In this report, we investigated the role of divalent cations in adiponectin assembly. Purified adiponectin 18mers, the largest HMW species, did not collapse to smaller oligomers after treatment with high concentrations of EDTA. However, treatment with EDTA or another chelator DTPA inhibited the oligomerization of 18mers from trimers in vitro. Zn(2+) specifically increased the formation of 18mers when compared with Cu(2+), Mg(2+), and Ca(2+). Distribution of adiponectin oligomers secreted from zinc chelator TPEN-treated rat adipocytes skewed toward increased proportions of hexamers and trimers. While we observed presence of zinc in adiponectin purified from calf serum, the role of zinc in disulfide bonding between oligomers was examined because the process is critical for 18mer assembly. Surprisingly, Zn(2+) inhibited disulfide bond formation early in the oligomerization process. We hypothesize that initial decreases in disulfide formation rates could allow adiponectin subunits to associate before becoming locked in fully oxidized conformations incapable of further oligomerization. These data demonstrate that zinc stimulates oligomerization of HMW adiponectin and possibly other disulfide-dependent protein assembly processes.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, MRB Diabetes Research, Tucson, AZ 85724, USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Briggs
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Giron
authors.forename: Karina
authors.initials: K
authors.lastname: Schnittker
authors.forename: Madeline V
authors.initials: MV
authors.lastname: Hart
authors.forename: Chad K
authors.initials: CK
authors.lastname: Park
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Hausrath
authors.forename: Tsu-Shuen
authors.initials: TS
authors.lastname: Tsao
mesh_terms: D052242:Adiponectin / Q000737:chemistry*; D000818:Animals; D004220:Disulfides / Q000737:chemistry*; D004229:Dithiothreitol / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D005029:Ethylenediamines / Q000494:pharmacology; D005260:Female; D006863:Hydrogen-Ion Concentration; D004369:Pentetic Acid / Q000494:pharmacology; D055503:Protein Multimerization; D051381:Rats; D011924:Rats, Zucker; D015032:Zinc / Q000494:pharmacology*
chemical_list: D052242:Adiponectin; D004220:Disulfides; D005029:Ethylenediamines; D004369:Pentetic Acid; D004492:Edetic Acid; D015032:Zinc; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine; D004229:Dithiothreitol