2A99

Crystal structure of recombinant chicken sulfite oxidase at resting state

2A99 の概要

• エントリーDOI: 10.2210/pdb2a99/pdb
• 関連するPDBエントリー: 2A9A 2A9B 2A9C 2A9D
• 分子名称: Sulfite Oxidase, CHLORIDE ION, PHOSPHONIC ACIDMONO-(2-AMINO-5,6-DIMERCAPTO-4-OXO-3,7,8A,9,10,10A-HEXAHYDRO-4H-8-OXA-1,3,9,10-TETRAAZA-ANTHRACEN-7-YLMETHYL)ESTER, ... (6 entities in total)
• 機能のキーワード: sulfite oxidase; molybdopterin; molybdenum, oxidoreductase
• 由来する生物種: Gallus gallus (chicken)
• 細胞内の位置: Mitochondrion intermembrane space: P07850
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 41252.66

構造登録者

Karakas, E.,Wilson, H.L.,Graf, T.N.,Xiang, S.,Jaramillo-Busquets, S.,Rajagopalan, K.V.,Kisker, C. (登録日: 2005-07-11, 公開日: 2005-08-02, 最終更新日: 2023-08-23)

主引用文献

Karakas, E.,Wilson, H.L.,Graf, T.N.,Xiang, S.,Jaramillo-Busquets, S.,Rajagopalan, K.V.,Kisker, C.
Structural insights into sulfite oxidase deficiency
J.Biol.Chem., 280:33506-33515, 2005

PubMed Abstract: Sulfite oxidase deficiency is a lethal genetic disease that results from defects either in the genes encoding proteins involved in molybdenum cofactor biosynthesis or in the sulfite oxidase gene itself. Several point mutations in the sulfite oxidase gene have been identified from patients suffering from this disease worldwide. Although detailed biochemical analyses have been carried out on these mutations, no structural data could be obtained because of problems in crystallizing recombinant human and rat sulfite oxidases and the failure to clone the chicken sulfite oxidase gene. We synthesized the gene for chicken sulfite oxidase de novo, working backward from the amino acid sequence of the native chicken liver enzyme by PCR amplification of a series of 72 overlapping primers. The recombinant protein displayed the characteristic absorption spectrum of sulfite oxidase and exhibited steady state and rapid kinetic parameters comparable with those of the tissue-derived enzyme. We solved the crystal structures of the wild type and the sulfite oxidase deficiency-causing R138Q (R160Q in humans) variant of recombinant chicken sulfite oxidase in the resting and sulfate-bound forms. Significant alterations in the substrate-binding pocket were detected in the structure of the mutant, and a comparison between the wild type and mutant protein revealed that the active site residue Arg-450 adopts different conformations in the presence and absence of bound sulfate. The size of the binding pocket is thereby considerably reduced, and its position relative to the cofactor is shifted, causing an increase in the distance of the sulfur atom of the bound sulfate to the molybdenum.

PubMed: 16048997
DOI: 10.1074/jbc.M505035200

実験手法

X-RAY DIFFRACTION (2.202 Å)