IMMUNOCHEMISTRY OF MAMMALIAN MITOCHONDRIAL RIBOSOMES

哺乳动物线粒体含有55S核糖体，其组成异常， 物理化学性质。拟议的研究将使用 哺乳动物蛋白质组成的免疫化学分析方法 线粒体核糖体，并阐明其生物合成、结构 哺乳动物几种蛋白质的组织和功能作用 线粒体核糖体。大规模制备线粒体核糖体 将被用来在兔子身上产生抗体，并 小鼠抗亚核糖体颗粒和蛋白质的单抗 亚核糖体颗粒的提取物。这些抗体将用于 关于生物合成、性质和进化的各种研究 哺乳动物的有丝分裂体蛋白。 针对牛mtr-蛋白的抗体将用于筛查人类或 牛在lambda gt11中的表达文库及克隆的DNA插入 获得的基因将用于筛选牛或人类基因组文库 相应的mtr-蛋白基因。克隆的cDNAs和基因组序列 将被分析以确定序列、结构和组织 哺乳动物有丝分裂体蛋白的基因。两种编码方式的比较 与成熟r-蛋白N端氨基酸序列的同源性 可能揭示存在于前体r-蛋白中的N末端转运肽。 人和牛特定mt基因编码序列的比较 R-蛋白基因应该揭示这些差异的性质和程度 基因的进化似乎比细胞质基因进化得更快 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.