title: Defective protein folding and intracellular retention of thyroglobulin-R19K mutant as a cause of human congenital goiter.
abstract: It has been suggested that a thyroglobulin (Tg)-R19K missense mutation may be a newly identified cause of human congenital goiter, which is surprising for this seemingly conservative substitution. Here, we have examined the intracellular fate of recombinant mutant Tg expressed in COS-7 cells. Incorporation of the R19K mutation largely blocked Tg secretion, and this mutant was approximately 90% degraded intracellularly over a 24-h period after synthesis. Before its degradation, the Tg-R19K mutant exhibited abnormally increased association with molecular chaperones BiP, calnexin, and protein disulfide isomerase, and was unable to undergo anterograde advance from the endoplasmic reticulum (ER) through the Golgi complex. Inhibitors of proteasomal proteolysis and ER mannosidase-I both prevented ER-associated degradation of the Tg-R19K mutant and increased its association with ER molecular chaperones. ER quality control around Tg residue 19 is not dependent upon charge but upon side-chain packing, because Tg-R19Q was efficiently secreted. Whereas a Tg mutant truncated after residue 174 folds sufficiently well to escape ER quality control, introduction of the R19K point mutation blocked its secretion. The data indicate that the R19K mutation induces local misfolding in the amino-terminal domain of Tg that has global effects on Tg transport and thyroid hormonogenesis.
authors.affiliation: Division of Metabolism, Endocrinology & Diabetes, University of Michigan Medical Center, Ann Arbor, MI 48109, USA.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Kim
authors.forename: Jaemin
authors.initials: J
authors.lastname: Lee
authors.forename: Piyanuch
authors.initials: P
authors.lastname: Jongsamak
authors.forename: Shekar
authors.initials: S
authors.lastname: Menon
authors.forename: Bailing
authors.initials: B
authors.lastname: Li
authors.forename: Shaikh A
authors.initials: SA
authors.lastname: Hossain
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Bae
authors.forename: Bhinyo
authors.initials: B
authors.lastname: Panijpan
authors.forename: Peter
authors.initials: P
authors.lastname: Arvan
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D019943:Amino Acid Substitution; D000818:Animals; D001120:Arginine / Q000235:genetics; D001692:Biological Transport; D015153:Blotting, Western; D019556:COS Cells; D037281:Calnexin / Q000378:metabolism; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D006042:Goiter / Q000235:genetics* / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D007976:Leupeptins / Q000494:pharmacology; D008239:Lysine / Q000235:genetics; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D017510:Protein Folding; D015398:Signal Transduction / Q000187:drug effects; D013954:Thyroglobulin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D000470:Alkaloids; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D007976:Leupeptins; D018832:Molecular Chaperones; C065629:kifunensine; D037281:Calnexin; D013954:Thyroglobulin; D001120:Arginine; D019704:Protein Disulfide-Isomerases; D008239:Lysine; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde