IMMUNOCHEMISTRY OF MAMMALIAN MITOCHONDRIAL RIBOSOMES

哺乳动物线粒体含有55 S核糖体的不寻常的组成， 物理化学性质 该研究将使用 免疫化学方法分析哺乳动物的蛋白质组成 线粒体核糖体，并阐明生物合成，结构 哺乳动物的几种蛋白质的组织和功能作用 线粒体核糖体 大规模制备的线粒体核糖体 牛肝脏中的蛋白质将用于在兔子中产生抗体， 抗核糖体颗粒和蛋白质的小鼠单克隆抗体 亚核糖体颗粒的提取物。 这些抗体将用于 对生物合成、性质和进化的各种研究 哺乳动物有丝分裂体蛋白。 抗牛mtr蛋白的抗体将用于筛选人或 λ gt 11中的牛cDNA表达文库和克隆的DNA插入片段 获得的基因组文库将用于筛选牛或人基因组文库， 相应的线粒体R蛋白基因。 克隆的cDNA和基因组序列 将进行分析，以确定序列，结构和组织 哺乳动物线粒体蛋白基因。 编码比较 序列与成熟r-蛋白的N-末端氨基酸序列 可以揭示前体r蛋白中存在的N-末端转运肽。 特定mt的人和牛编码序列的比较 r-蛋白基因应该揭示这些差异的性质和程度。 基因，这似乎是进化得更快，比那些细胞质 R蛋白

Mammalian mitochondria contain 55S ribosomes of unusual composition and physical-chemical properties. The proposed research will use immunochemical methods to analyze the protein composition of mammalian mitochondrial ribosomes, and to elucidate the biosynthesis, structural organization, and functional roles of several of the proteins of mammalian mitochondrial ribosomes. Mitochondrial ribosomes prepared on a large scale from bovine liver will be used to produce antibodies in rabbits and monoclonal antibodies in mice against subribosomal particles and protein extracts of subribosomal particles. These antibodies will be used in a variety of studies on the biosynthesis, properties and evolution of mammalian mitoribosomal proteins. Antibodies against bovine mt r-proteins will be used to screen human or bovine cDNA expression libraries in lambda gt11, and the cloned DNA inserts obtained will be used to screen bovine or human genomic libraries for the corresponding mt r-protein genes. The cloned cDNA and genomic sequences will be analyzed to determine the sequence, structure and organization of genes for mammalian mitoribosomal proteins. Comparison of the coding sequence with the N-terminal amino acid sequence of the mature r-proteins may reveal N-terminal transit peptides present in precursor r-proteins. Comparison of the human and bovine coding sequences for particular mt r-protein genes should reveal the nature and extent of divergence of these genes, which appear to be evolving more rapidly than those for cytoplasmic r-proteins.