Histidine residue at position 226 is critical for iodide uptake activity of human sodium/iodide symporter.
The sodium/iodide symporter (SLC5A5; also known as NIS), a transmembrane glycoprotein principally in the thyroid gland, is responsible for the accumulation of iodide necessary for thyroid hormones. Our previous study indicated that a novel exon 6 deletion (residues 233-280) in SLC5A5 loses the iodide uptake activity. Herein we characterized the role of His-226 in iodide transport of SLC5A5. His-226, a highly conserved extracellular residue among SLC5A5 homologs, was replaced with alanine, aspartic acid, glutamic acid, or lysine. All the SLC5A5 mutants were expressed normally in the cells and targeted correctly to the plasma membrane. However, all of the mutants displayed severe defects in iodide uptake, suggesting that His-226 was critical for iodide uptake. Kinetic analysis further showed that mutation at His-226 led to a dramatic decrease in V(max). These findings suggested that the decreased levels of iodide uptake activity of SLC5A5 mutants resulted from lower catalytic rates. In conclusion, our data first identified the involvement of extracellular charged amino acid residue in the iodide uptake ability of SLC5A5.
Wu
Shih-Lu
SL
Department of Biochemistry, China Medical University, 91 Hsueh-Shih Road, Taichung 40402, Taiwan, ROC.
Ho
Tin-Yun
TY
Liang
Ji-An
JA
Hsiang
Chien-Yun
CY
D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D003198:Computer Simulation; D005455:Fluorescent Antibody Technique; D006639:Histidine / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007454:Iodides / Q000378:metabolism*; D007700:Kinetics; D018613:Microscopy, Confocal; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D027981:Symporters / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000099209:Sodium-Iodide Symporters
D006639:Histidine; D007454:Iodides; D027981:Symporters; D000099209:Sodium-Iodide Symporters