Oxidation of glyceraidehyde-3-phosphate dehydrogenase enhances its binding to nucleic acids
Oxidation of glyceraidehyde-3-phosphate dehydrogenase enhances its binding to nucleic acids
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DOI：
10.1016/s0006-291x(03)01222-1
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发表时间：
2003-08-01
影响因子：
3.1
通讯作者：
Muronetz, VI
中科院分区：
文献类型：
--
作者：
Arutyunova, EI;Danshina, PV;Muronetz, VI
甘油醛-3-磷酸脱氢酶（GAPDH）是一种活性与其酶功能相差甚远的蛋白质。在这里，我们表明，氧化的SH-基团的GAPDH的活性位点增强其与总的转移RNA或总DNA的结合。GAPDH的辅因子NAD和NADH均能抑制GAPDH与RNA（DNA）的相互作用，但在氧化型GAPDH中，NAD的抑制作用不如NADH。GAPDH的氧化强烈降低其对NAD的亲和力，但不降低对NADH的亲和力。固定化的GAPDH四聚体在与总RNA而不是DNA孵育期间解离成二聚体。用特异于GAPDH的二聚体、单体或变性形式的单克隆抗体对HeLa细胞进行染色，揭示了非天然形式的GAPDH在细胞核中的浓缩。GAPDH的氧化在酶的四级结构的调节和在其与核酸的相互作用的作用进行了讨论。(C)2003年爱思唯尔公司All rights reserved.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a protein with various activities far from its enzymatic function. Here, we showed that the oxidation of SH-groups of the active site of GAPDH enhanced its binding with total transfer RNA or with total DNA. Both NAD and NADH-the cofactors of GAPDH-inhibited the GAPDH-RNA (DNA) interaction, though NAD was much less effective than NADH in the case of oxidized GAPDH. Oxidation of GAPDH strongly decreased its affinity to NAD but not to NADH. Immobilized tetramers of GAPDH dissociated into dimers during the incubation with total RNA but not DNA. The staining of HeLa cells with monoclonal antibodies specific to dimers, monomers or the denatured form of GAPDH revealed the condensation of non-native forms of GAPDH in the nucleus. The role of oxidation of GAPDH in the regulation of the quaternary structure of the enzyme and in its interaction with nucleic acids is discussed. (C) 2003 Elsevier Inc. All rights reserved.