PROTEINS IN LEAD-INDUCED NUCLEAR INCLUSION BODIES

铅中毒的临床影响是众所周知的，但有 对低水平铅暴露可能产生的影响的担忧日益加剧。 这项提议针对的是一种核蛋白，它是 摄入的铅。这种名为p32/6.3的蛋白质是铅的丰富成分。 在肾脏中诱导出核内包涵体。在正常成年人中是这样的 在中枢神经系统神经元中含量最高，增加的原因是 突触成熟。长期目标是了解铅的影响 在脑以及肾脏和其他细胞中的p32/6.3上 通常是低丰度的成分，而且要进一步理解 这些影响的后果。一种单抗将被用于检测 几种铅暴露模型的p32/6.3池大小：培养的脑 微血管内皮细胞，大脑，小脑，脑微血管， 成人和母亲铅暴露的新生儿的肾脏；以及 EDTA络合治疗后的成人肾脏。使用来自 部分肽序列、聚合酶链式反应和基因克隆 将被用来完成序列；这可能确认与 CAMP结合蛋白。利用cdna序列，融合蛋白将被用作 抗原；产生的抗体将用于执行 上述模型中的免疫细胞化学。一个相关的项目将 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.