Mapping of Ion and Substrate Binding Sites in Human Sodium Iodide Symporter (hNIS)

被引:4
作者
Zhekova, Hristina R. [1 ]
Sakuma, Toshie [2 ,3 ]
Johnson, Ryan [2 ]
Concilio, Susanna C. [3 ,4 ]
Lech, Patrycja J. [2 ]
Zdravkovic, Igor [1 ]
Damergi, Mirna [1 ]
Suksanpaisan, Lukkana [2 ]
Peng, Kah-Whye [2 ,3 ]
Russell, Stephen J. [2 ,3 ]
Noskov, Sergei [1 ]
机构
[1] Univ Calgary, Ctr Mol Simulat, Dept Biol Sci, Calgary, AB T2N 1N4, Canada
[2] Imanis Life Sci, Rochester, MN 55901 USA
[3] Mayo Clin, Dept Mol Med, Rochester, MN 55902 USA
[4] Mayo Clin, Grad Sch Biomed Sci, Rochester, MN 55902 USA
基金
加拿大自然科学与工程研究理事会;
关键词
THYROID NA+/I-SYMPORTER; NA+/PROLINE TRANSPORTER PUTP; TRANSMEMBRANE DOMAIN-IX; AMINO-ACID RESIDUE; SODIUM/IODIDE SYMPORTER; ESCHERICHIA-COLI; ALTERNATING-ACCESS; MEMBRANE-TRANSPORT; CRYSTAL-STRUCTURE; CONFORMATIONAL DYNAMICS;
D O I
10.1021/acs.jcim.9b01114
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
The human sodium iodide symporter (hNIS) is a theranostic reporter gene which concentrates several clinically approved SPECT and PET radiotracers and plays an essential role for the synthesis of thyroid hormones as an iodide transporter in the thyroid gland. Development of hNIS mutants which could enhance translocation of the desired imaging ions is currently underway. Unfortunately, it is hindered by lack of understanding of the 3D organization of hNIS and its relation to anion transport. There are no known crystal structures of hNIS in any of its conformational states. Homology modeling can be very effective in such situations; however, the low sequence identity between hNIS and relevant secondary transporters with available experimental structures makes the choice of a template and the generation of 3D models nontrivial. Here, we report a combined application of homology modeling and molecular dynamics refining of the hNIS structure in its semioccluded state. The modeling was based on templates from the LeuT-fold protein family and was done with emphasis on the refinement of the substrate-ion binding pocket. The consensus model developed in this work is compared to available biophysical and biochemical experimental data for a number of different LeuT-fold proteins. Some functionally important residues contributing to the formation of putative binding sites and permeation pathways for the cotransported Na+ ions and I- substrate were identified. The model predictions were experimentally tested by generation of mutant versions of hNIS and measurement of relative (to WT hNIS) I-125(-) uptake of 35 hNIS variants.
引用
收藏
页码:1652 / 1665
页数:14
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