PROTEINS IN LEAD-INDUCED NUCLEAR INCLUSION BODIES

铅中毒的临床影响是众所周知的，但 人们越来越关注低水平铅暴露的可能影响。 这项提议针对的是一种核蛋白， 摄入了铅 蛋白质p32/6.3是铅的丰富成分， 在肾脏中诱导核内包涵体。 在正常成年人中， 最丰富的中枢神经系统神经元，增加发生在 突触成熟。 长期目标是了解铅的影响 p32/6.3在大脑以及肾脏和其他细胞中， 通常是一个低丰度的组成部分，并进一步了解， 这些影响的后果。 将使用单克隆抗体测定 几种铅暴露模型中p32/6.3池的大小：培养的脑 微血管内皮细胞;大脑，小脑，脑微血管， 以及母亲接触铅的成人和新生儿的肾脏;以及 EDTA螯合治疗后的成人肾脏。 使用来自 部分肽段序列、聚合酶链反应和cDNA克隆 将用于完成序列;这可以证实与一个人的同源性。 cAMP结合蛋白。 使用cDNA序列，融合蛋白将用作 抗原;产生的抗体将用于执行 免疫细胞化学在上述模型中。 相关项目将 研究铅诱导蛋白的诱导机制。

The clinical effects of lead intoxication are well known, but there is increasing concern over the possible effects of low level lead exposure. This proposal is directed at a nuclear protein which is a target for ingested lead. The protein, p32/6.3, is an abundant component of lead- induced intranuclear inclusion bodies in kidney. In normal adults it is most abundant in central nervous system neurons, the increase occurring as synapses mature. The long term objectives are to understand lead's effects on p32/6.3 in brain as well as in kidney and other cells where it is normally a low-abundance component, and further, to understand the consequences of these effects. A monoclonal antibody will be used to assay p32/6.3 pool size in several lead exposed models: cultured brain microvessel endothelial cells; cerebrum, cerebellum, brain microvessels, and kidney of adults and of neonates whose mothers are lead exposed; and kidney of adults after EDTA chelation therapy. Using information from a partial peptide sequence, the polymerase chain reaction and cDNA cloning will be used to complete the sequence; this may confirm homology with a cAMP-binding protein. Using cDNA sequence, fusion proteins will be used as antigens; the resultant antibodies will be used to perform immunocytochemistry in the models described above. A related project will study the mechanism of induction of a lead-induced protein.