Characterization of the thyroid Na+/I- symporter with an anti-COOH terminus antibody

Orlie Levy, Ge Dai, Claudia Riedel, Christopher S. Ginter, Elliot M. Paul, Adam N. Lebowitz, Nancy Carrasco

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

191 Citas (Scopus)


The Na+/I- symporter (NIS) is the plasma membrane protein that catalyzes active I- transport in the thyroid, the first step in thyroid hormone biogenesis. The cDNA encoding NIS was recently cloned in our laboratory and a secondary structure model proposed, suggesting that NIS is an intrinsic membrane protein (618 amino acids; ≃65.2 kDa predicted molecular mass) with 12 putative transmembrane domains. Here we report the generation of a site-directed polyclonal anti-COOH terminus NIS antibody (Ab) that immunoreacts with a ≃87 kDa-polypeptide present in membrane fractions from a rat thyroid cell line (FRTL-5). The model-predicted cytosolic-side location of the COOH terminus was confirmed by indirect immunofluorescence experiments using anti-COOH terminus NIS Ab in permeabilized FRTL-5 cells. Immunoreactivity was competitively blocked by the presence of excess synthetic peptide. Treatment of membrane fractions from FRTL-5 cells, Xenopus laevis oocytes, and COS cells expressing NIS with peptidyl N-glycanase F converted the ≃87 kDa-polypeptide into a ≃50 kDa-species, the same relative molecular weight exhibited by NIS expressed in E. coli. Anti-NIS Ab immunoprecipitated both the NIS precursor molecule (≃56 kDa) and the mature ≃87 kDa form. Furthermore, a direct correlation between circulating levels of thyroid-stimulating hormone and NIS expression in vivo was demonstrated.

Idioma originalInglés
Páginas (desde-hasta)5568-5573
Número de páginas6
PublicaciónProceedings of the National Academy of Sciences of the United States of America
EstadoPublicada - 27 may 1997

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