Identification and selective precipitation of human aldehyde dehydrogenase isozymes using antibodies raised to horse liver aldehyde dehydrogenase isozymes.
Identification and selective precipitation of human aldehyde dehydrogenase isozymes using antibodies raised to horse liver aldehyde dehydrogenase isozymes.
复制标题
使用针对马肝醛脱氢酶同工酶产生的抗体来鉴定和选择性沉淀人醛脱氢酶同工酶。
DOI：
10.1111/j.1530-0277.1986.tb05098.x
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Weiner,H
中科院分区：
文献类型：
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作者：
McMichael,M;Hellström-Lindahl,E;Weiner,H
在免疫印迹实验中识别来自人肝匀浆的醛脱氢酶（ALDH），并通过针对马肝线粒体和胞质ALDH同工酶的抗体在Ouchterlony双扩散凝胶中沉淀。针对马肝细胞质Rc ALDH（α HC）产生的抗体已被证明对细胞质ALDH同工酶具有特异性，而针对马肝线粒体ALDH（α HM）产生的抗体沉淀线粒体和细胞质ALDH同工酶。通过与α HC预孵育，可以选择性地从α高加索人肝脏样品的匀浆中去除胞浆ALDH;然后在双扩散凝胶中通过aHM沉淀剩余的线粒体酶。用来自东方人的肝脏样本重复实验，由于没有发现活性线粒体ALDH，因此推测为酒精敏感。α HM从不含ALDH的细胞溶质样本中沉淀出相对无活性的线粒体酶，证实了先前关于酒精敏感东方人组织中存在线粒体ALDH的报道。免疫印迹实验的结果证实了来自酒精敏感东方人肝脏的胞质和线粒体ALDH在电泳中的共迁移。这里报告的结果，连同以前的观察，表明马肝ALDH同工酶的抗体，可用于确定ALDH同工酶在各种人体组织中的亚细胞位置，包括冷冻组织样品，不适合亚细胞分级。
Aldehyde dehydrogenase (ALDH) enzymes from human liver homogenates were recognized in immunoblotting experiments and precipitated in Ouchterlony double diffusion gels by antibodies raised to the horse liver mitochondrial and cvtosolic ALDH isozymes. The antibody raised to the cytosoRc horse liver ALDH (αHC) has been shown to be specific for cytosolic ALDH isozymes, while the antibody raised to the horse liver mitochondrial ALDH (αHM) precipitated both mitochondrial and cytosolic ALDH isozymes. It was possible to selectively remove the cytosolic ALDH from a homogenate of a liver sample from α Caucasian by preincubation with αHC; the remaining mitochondrial enzyme was then precipitated by aHM in double diffusion gels. The experiments were repeated with a liver sample from an Oriental, presumed to have been alcohol sensitive since no active mitochondrial ALDH was found. The precipitation of a relatively inactive mitochondrial enzyme by αHM from a cytosolic ALDH‐free sample confirmed previous reports of the existence of a mitochondrial ALDH in tissue from an alcohol‐sensitive Oriental. The results of immunoblotting experiments confirm the co‐migration, in electrophoresis, of the cytosolic and mitochondrial ALDHs from the liver of an alcohol‐sensitive Oriental. The results reported here, together with previous observations, indicate that the antibodies raised to horse liver ALDH isozymes can be used to determine the subcellular location of ALDH isozymes in various human tissues, including frozen tissue samples which are not amenable to subcellular fractionation.