Traoré Daouda A K, El Ghazouani Abdelnasser, Jacquamet Lilian, Borel Franck, Ferrer Jean-Luc, Lascoux David, Ravanat Jean-Luc, Jaquinod Michel, Blondin Geneviève, Caux-Thang Christelle, Duarte Victor, Latour Jean-Marc
Commissariat à l'Energie Atomique, Institut de Recherches en Technologies et Sciences pour le Vivant, Laboratoire de Chimie et Biologie des Métaux, CEA-Grenoble, 17 avenue des Martyrs, 38054 Grenoble Cedex 9, France.
Nat Chem Biol. 2009 Jan;5(1):53-9. doi: 10.1038/nchembio.133. Epub 2008 Dec 14.
In Bacillus subtilis, PerR is a metal-dependent sensor of hydrogen peroxide. PerR is a dimeric zinc protein with a regulatory site that coordinates either Fe(2+) (PerR-Zn-Fe) or Mn(2+) (PerR-Zn-Mn). Though most of the peroxide sensors use cysteines to detect H(2)O(2), it has been shown that reaction of PerR-Zn-Fe with H(2)O(2) leads to the oxidation of one histidine residue. Oxidation of PerR leads to the incorporation of one oxygen atom into His37 or His91. This study presents the crystal structure of the oxidized PerR protein (PerR-Zn-ox), which clearly shows a 2-oxo-histidine residue in position 37. Formation of 2-oxo-histidine is demonstrated and quantified by HPLC-MS/MS. EPR experiments indicate that PerR-Zn-H37ox retains a significant affinity for the regulatory metal, whereas PerR-Zn-H91ox shows a considerably reduced affinity for the metal ion. In spite of these major differences in terms of metal binding affinity, oxidation of His37 and/or His91 in PerR prevents DNA binding.
在枯草芽孢杆菌中，PerR是一种依赖金属的过氧化氢传感器。PerR是一种二聚体锌蛋白，具有一个可与Fe(2+)（PerR-Zn-Fe）或Mn(2+)（PerR-Zn-Mn）配位的调节位点。尽管大多数过氧化物传感器利用半胱氨酸来检测H(2)O(2)，但研究表明，PerR-Zn-Fe与H(2)O(2)反应会导致一个组氨酸残基被氧化。PerR的氧化会使一个氧原子掺入His37或His91中。本研究展示了氧化型PerR蛋白（PerR-Zn-ox）的晶体结构，该结构清楚地显示在37位有一个2-氧代组氨酸残基。通过HPLC-MS/MS证明并定量了2-氧代组氨酸的形成。电子顺磁共振实验表明，PerR-Zn-H37ox对调节金属仍具有显著亲和力，而PerR-Zn-H91ox对金属离子的亲和力则大幅降低。尽管在金属结合亲和力方面存在这些主要差异，但PerR中His37和/或His91的氧化会阻止DNA结合。
