title: Secretion of the adipocyte-specific secretory protein adiponectin critically depends on thiol-mediated protein retention.
abstract: Adiponectin is a secretory protein abundantly secreted from adipocytes. It assembles into a number of different higher-order complexes. Adipocytes maintain tight control over circulating plasma levels, suggesting the existence of a complex, highly regulated biosynthetic pathway. However, the critical mediators of adiponectin maturation within the secretory pathway have not been elucidated. Previously, we found that a significant portion of de novo-synthesized adiponectin is not secreted and retained in adipocytes. Here, we show that there is an abundant pool of properly folded adiponectin in the secretory pathway that is retained through thiol-mediated retention, as judged by the release of adiponectin in response to treatment of adipocytes with reducing agents. Adiponectin is covalently bound to the ER chaperone ERp44. An adiponectin mutant lacking cysteine 39 fails to stably interact with ERp44, demonstrating that this residue is the primary site mediating the covalent interaction. Another ER chaperone, Ero1-Lalpha, plays a critical role in the release of adiponectin from ERp44. Levels of both of these proteins are highly regulated in adipocytes and are influenced by the metabolic state of the cell. While less critical for the secretion of trimers, these chaperones play a major role in the assembly of higher-order adiponectin complexes. Our data highlight the importance of posttranslational events controlling adiponectin levels and the release of adiponectin from adipocytes. One mechanism for increasing circulating levels of specific adiponectin complexes by peroxisome proliferator-activated receptor gamma agonists may be selective upregulation of rate-limiting chaperones.
authors.affiliation: Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
authors.forename: Zhao V
authors.initials: ZV
authors.lastname: Wang
authors.forename: Todd D
authors.initials: TD
authors.lastname: Schraw
authors.forename: Ja-Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Tayeba
authors.initials: T
authors.lastname: Khan
authors.forename: Michael W
authors.initials: MW
authors.lastname: Rajala
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Philipp E
authors.initials: PE
authors.lastname: Scherer
mesh_terms: D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D052242:Adiponectin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003513:Cycloheximide / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D010088:Oxidoreductases; D047495:PPAR gamma / Q000819:agonists; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D034622:RNA Interference; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D052242:Adiponectin; C520556:ERp44 protein, mouse; D006023:Glycoproteins; D007328:Insulin; D008565:Membrane Proteins; D018832:Molecular Chaperones; D047495:PPAR gamma; D011500:Protein Synthesis Inhibitors; D013438:Sulfhydryl Compounds; D003513:Cycloheximide; C491864:Ero1l protein, mouse; D010088:Oxidoreductases