3TO0

Crystal structure of Mus musculus iodotyrosine deiodinase (IYD) C217A, C239A bound to FMN

3TO0 の概要

• エントリーDOI: 10.2210/pdb3to0/pdb
• 関連するPDBエントリー: 3GB5 3GFD 3GH8 3TNZ
• 分子名称: Iodotyrosine deiodinase 1, FLAVIN MONONUCLEOTIDE, PHOSPHATE ION, ... (5 entities in total)
• 機能のキーワード: oxidoreductase, flavoprotein, membrane, transmembrane, dehalogenase, iodide salvage, mono-iodotyrosine, nadp
• 由来する生物種: Mus musculus (mouse)
• 細胞内の位置: Membrane; Single-pass membrane protein (By similarity): Q9DCX8
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 61067.26

構造登録者

Buss, J.M.,McTamney, P.M.,Rokita, S.E. (登録日: 2011-09-02, 公開日: 2012-02-29, 最終更新日: 2023-09-13)

主引用文献

Buss, J.M.,McTamney, P.M.,Rokita, S.E.
Expression of a soluble form of iodotyrosine deiodinase for active site characterization by engineering the native membrane protein from Mus musculus.
Protein Sci., 21:351-361, 2012

PubMed Abstract: Reductive deiodination is critical for thyroid function and represents an unusual exception to the more common oxidative and hydrolytic mechanisms of dehalogenation in mammals. Studies on the reductive processes have been limited by a lack of convenient methods for heterologous expression of the appropriate proteins in large scale. The enzyme responsible for iodide salvage in the thyroid, iodotyrosine deodinase, is now readily generated after engineering its gene from Mus musculus. High expression of a truncated derivative lacking the membrane domain at its N-terminal was observed in Sf9 cells, whereas expression in Pichia pastoris remained low despite codon optimization. Ultimately, the desired expression in Escherichia coli was achieved after replacing the two conserved Cys residues of the deiodinase with Ala and fusing the resulting protein to thioredoxin. This final construct provided abundant enzyme for crystallography and mutagenesis. Utility of the E. coli system was demonstrated by examining a set of active site residues critical for binding to the zwitterionic portion of substrate.

PubMed: 22238141
DOI: 10.1002/pro.2020

実験手法

X-RAY DIFFRACTION (2.655 Å)