Receptor-Mediated Endocytosis experimental pathway.pptVIP

Receptor-Mediated Endocytosis experimental pathway 1.The process of receptor binding and internalization 2.The LDL receptor 3.Internalization signals Technique — electron microscope  Michael Brown Joseph Goldstein (University of Texas Medical School in Dallas) Richard Anderson Experimental pathway 1.The group incubated fibroblasts（成纤维细胞） from normal and FH subjects with LDL that had been covalently linked（共价结合） to the iron-containing protein ferritin（铁蛋白） .这个研究小组从正常的和带有LDL的FH中慢慢提取成纤维细胞，LDL被共价的连接于铁蛋白中。 3.A temperature of 4℃ Ligands（配体） can bind to the cell surface but cannot be internalized（内化）, LDL-ferritin particles were seen to be bound to the cell surface. 当在4℃时，配体可以与细胞表面结合，但不能被内化吸收，LDL—铁蛋白微粒被发现结合于细胞表面。 Found: 1、The LDL particles were not randomly scattered over the cell surface but were localized to some segments of plasma membrane —— coated pits. The results supported the proposal that the mutant FH allele encoded receptor that was unable to bind LDL. 这结果显示了突变的FH等位基因翻译成受体，而这个受体并没有能力结合LDL。 Subsequent electron microscopic studies on the internalization of LDL-ferritin revealed the endocytic pathway by which these lipoprotein particles were internalized 后来用电子显微镜对LDL—铁蛋白的内化作用进行研究，显示出胞吞作用的路径是这些脂蛋白的内化。 Postulated that … The rapid internalization of receptor-bound LDL is strictly dependent on the localization of LDL receptors in coated pits. 基于这些结论，这个小组假定受体结合LDL的内化作用是在有被小窝中LDL受体的某部位密切相关的。 J.D. mutation At about this time, an LDL receptor with a different type of mutation was discovered. LDL receptors bearing this new defect bound normal amounts of radioactively labeled LDL, yet the receptor-bound lipoprotein failed to internalized and consequently was not delivered to cytoplasmic lysosomes for processing. LDL受体是一种多机能蛋白，由839个氨基酸组成的36面体结构蛋白，分子量约115kD。由五种不同的区域构成，各区域有其独特的功能 。 J.D. mutation Analysis of the J.D. mutant receptor revealed that the protein contained a single amino acid substitution: a tyrosine(酪氨