Glyceraldehyde 3-Phosphate Dehydrogenase Is Unlikely to Mediate Hydrogen Peroxide Signaling: Studies with a Novel Anti-Dimedone Sulfenic Acid Antibody
Glyceraldehyde 3-Phosphate Dehydrogenase Is Unlikely to Mediate Hydrogen Peroxide Signaling: Studies with a Novel Anti-Dimedone Sulfenic Acid Antibody
复制标题
DOI：
10.1089/ars.2010.3149
复制
发表时间：
2011-01-01
影响因子：
6.6
通讯作者：
Eaton, Philip
中科院分区：
文献类型：
--
作者：
Maller, Claire;Schroeder, Ewald;Eaton, Philip
蛋白质磺酸(SOHs)是氧化还原活性蛋白质与过氧化氢分子相互作用的主要氧化产物。我们开发了一种新的抗体试剂，可以检测由双酮衍生的蛋白SOH。使用这种新的抗体，我们发现在基础条件下，甘油醛3-磷酸脱氢酶(GAPDH)是在基础条件下存在于分离的大鼠心室肌细胞中的主要蛋白质亚硫酸盐。在过氧化氢(H_2O_2)的氧化应激过程中，GAPDHSOH标记丢失，但随后出现一些次级二酮反应蛋白硫酸盐。随着磺酸盐标记的丢失，GAPDH的半胱氨酸-149亚磺酸/磺酸氧化态出现。这种高氧化的GAPDH既与糖酵解的抑制有关，也与其还原过氧化氢的能力有关。我们研究了GAPDH的失活是否导致了与其过度氧化一致的次生蛋白硫酸盐的产生。选择性的GAPDH抑制剂柯宁酸(通过在Cys-149处形成共价加合物发挥作用)完全阻止了基础SOH标记以及随后的过氧化氢诱导的过氧化。然而，康宁酸介导的GAPDH的单独抑制不能诱导细胞内H_2O_2或次生蛋白硫酸盐的形成，也不能加强它们由过氧化氢诱导的形成。总体而言，GAPDH似乎具有过氧化物酶样属性，但它的抑制未能影响涉及次级蛋白质亚硫化的下游氧化剂信号。抗氧化剂。氧化还原信号。14，49-60。
Protein sulfenic acids (SOHs) are the principal oxidation products formed when redox active proteins interact with peroxide molecules. We have developed a new antibody reagent that detects protein SOHs derivatized with dimedone. Using this new antibody, we found that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is the predominant protein sulfenate present in isolated rat ventricular myocytes under basal conditions. During oxidative stress with hydrogen peroxide (H2O2), GAPDH SOH labeling is lost, but a number of secondary dimedone-reactive protein sulfenates then appear. As the sulfenate labeling is lost, the Cys-149 sulfinic/sulfonic acid oxidation states of GAPDH appear. This hyperoxidized GAPDH is associated with both the inhibition of glycolysis and its ability to reduce H2O2. We examined whether inactivation of GAPDH was causative in the generation of secondary protein sulfenates that coincide with its hyperoxidation. The selective GAPDH inhibitor koningic acid (which functions by forming a covalent adduct at Cys-149) fully prevented basal SOH labeling, as well as subsequent peroxide-induced hyperoxidation. However, koningic acid-mediated inhibition of GAPDH alone did not induce the formation of intracellular H2O2 or secondary protein sulfenates and also failed to potentiate their peroxide-induced formation. Overall, GAPDH appears to have peroxidase-like properties, but its inhibition failed to impact on downstream oxidant signaling involving secondary protein sulfenation. Antioxid. Redox Signal. 14, 49-60.