Asn441 plays a key role in folding and function of the Na +/I- symporter (NIS)

Wenjing Li, Juan Pablo Nicola, Mario L Amzel, Nancy Carrasco

Research output: Contribution to journalArticle

Abstract

The Na+/I-symporter (NIS) is a plasma membrane glycoprotein that mediates active I-transport in the thyroid, the first step in the biosynthesis of the iodine-containing thyroid hormones T3 and T4. Several NIS mutants have been identified as a cause of congenital I-transport defect (ITD), and their investigation has yielded valuable mechanistic information on NIS. Here we report a thorough characterization of the ITD-causing NIS mutation in which the sixth intracellular loop residues 439-443 are missing. This mutant protein was intracellularly retained, incompletely glycosylated, and intrinsically inactive. Engineering 5 Ala at positions 439-443 partially recovered cell surface targeting and activity (15%). Strikingly, NIS with the sequence 439-AANAA-443, in which Asn was restored at position 441, was targeted to the plasma membrane and exhibited 95% the transport activity of WT NIS. Based on our NIS homology model, we propose that the side chain of N441, a residue conserved throughout most of the SLC5 family, interacts with the main chain amino group of G444, capping the α-helix of transmembrane segment XII and thus stabilizing the structure of the molecule. Our data provide insight into a critical interhelical interaction required for NIS folding and activity.

Original languageEnglish (US)
Pages (from-to)3229-3238
Number of pages10
JournalFASEB Journal
Volume27
Issue number8
DOIs
StatePublished - Aug 2013

Fingerprint

Symporters
Cell membranes
Cell Membrane
Defects
Active Biological Transport
Biosynthesis
Membrane Glycoproteins
Triiodothyronine
Mutant Proteins
Thyroxine
Iodine
Thyroid Gland
Mutation
Molecules

Keywords

  • Congenital hypothyroidism
  • Helix capping
  • Interhelical interaction
  • Plasma membrane targeting
  • Protein folding

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology
  • Medicine(all)

Cite this

Asn441 plays a key role in folding and function of the Na +/I- symporter (NIS). / Li, Wenjing; Nicola, Juan Pablo; Amzel, Mario L; Carrasco, Nancy.

In: FASEB Journal, Vol. 27, No. 8, 08.2013, p. 3229-3238.

Research output: Contribution to journalArticle

Li, Wenjing ; Nicola, Juan Pablo ; Amzel, Mario L ; Carrasco, Nancy. / Asn441 plays a key role in folding and function of the Na +/I- symporter (NIS). In: FASEB Journal. 2013 ; Vol. 27, No. 8. pp. 3229-3238.
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