Development of a peptide antibody specific to human glutathione S-transferase alpha 4-4 (hGSTA4-4) reveals preferential localization in human liver mitochondria.
Development of a peptide antibody specific to human glutathione S-transferase alpha 4-4 (hGSTA4-4) reveals preferential localization in human liver mitochondria.
复制标题
人谷胱甘肽 S-转移酶 α 4-4 (hGSTA4-4) 特异性肽抗体的开发揭示了人肝线粒体中的优先定位。
DOI：
10.1006/abbi.2001.2352
复制
发表时间：
2001
影响因子：
3.9
通讯作者：
Gallagher,EP
中科院分区：
文献类型：
--
作者：
Gardner,JL;Gallagher,EP
在膜脂过氧化过程中产生的反应性细胞产物被认为是多种退行性疾病和衰老的病原体。尤其是4-羟基-2-烯醛(4HNE)是脂质过氧化过程中产生最多的物质之一。在人类和啮齿动物中，谷胱甘肽S转移酶的α4亚类(mGSTA4-4、rGSTA4-4、hGST-5.8和hGSTA4-4)对4HNE和其他羟基烯具有独特的高谷胱甘肽结合活性。在人体肝脏中，hGSTA4-4介导的4HNE结合似乎代表了4HNE解毒的高亲和力途径。在本研究中，我们研制了一种针对hGSTA4-4的高度特异性的多克隆抗体。人肝亚细胞组分的Western blotting分析和N-末端测序表明，hGSTA4-4定位于线粒体组分，但未在胞质组分中检测到。我们的结果提供了证据，证明在成人肝脏中，hGSTA4-4特异性地定位于线粒体，明显排除胞浆。HGSTA4-4靶向线粒体对于与有氧呼吸引起的氧化应激相关的退行性疾病具有潜在的意义。
The reactive cellular products generated during the peroxidation of membrane lipids have been implicated as causative agents in a variety of degenerative diseases and aging. In particular, 4-hydroxynon-2-enal (4HNE) is among the most of the produced during lipid peroxidation. In humans and rodent species, the alpha 4 subclass of glutathione S-transferases (mGSTA4-4, rGSTA4-4, hGST-5.8, and hGSTA4-4) exhibits uniquely high glutathione conjugation activity toward 4HNE and other hydroxyalkenals. In human liver, hGSTA4-4-mediated 4HNE conjugation appears to represent the high-affinity pathway for 4HNE detoxification. In the present study, a highly specific polyclonal antibody was developed against hGSTA4-4. Western blotting analysis of human liver subcellular fractions as well as N-terminal sequencing revealed that hGSTA4-4 was localized to mitochondrial fractions, but was not detected in cytosolic fractions. Our results provide evidence that in adult liver, hGSTA4-4 is specifically targeted to the mitochondrion to the apparent exclusion of the cytosol. Targeting of hGSTA4-4 to the mitochondrion holds implications for degenerative diseases associated with oxidative stress that arise from aerobic respiration.