生化及分子生物学原理及技术课蛋白质相互作用及核酸相互作用.ppt

* For example, signals from the exterior of a cell are mediated to the inside of that cell by protein-protein interactions of the signaling molecules In protein complex, members are linked by non-covalent interactions, they often activate or inhibit other members. a protein may be carrying another protein, for example, from cytoplasm to nucleus or vice versa in the case of the nuclear pore importins, a type of protein that moves other protein molecules into the nucleus by binding to a specific recognition sequence protein kinase will add a phosphate to a target protein * * Pioneered by Fields and Song in 1989 尽管真核生物体可以表达数万种不同的蛋白质，但对应的结构域、结构花样与折叠类型的数量却少得多。一种合理的解释是，这是进化的结果；因为基因或基因的一部分可以在基因组内被加倍或移动。也就是说，通过基因重组，一个结构域可以从相应蛋白质A移动到本不具有此结构域的蛋白质B上，而其发生的进化驱动力可能是由于该结构域对应的生物学功能趋向于被蛋白质B所利用。 * * * Chip Fabrication, Probing and Detection :Technical Issues High-throughput fusion protein purification Printing chips Suitable surface chemistry for attachment of proteins and retaining integrity, orientation, structure, activity Cross-contamination, Spot size, comets etc. Detection Sensitive-specific probe with Retain signal during washing Low background, high signal/noise * Design of protein chips Chip probed with ?-GST antibody and signals detected after Cy5-conjugated IgG 12,938 data points Each spot corresponds to ~30 fg-~50 pg protein * Analysis of the Yeast Proteome Chip Protein –Protein interactions 1° Ab against target protein domain 1° Ab against interacting partner protein Biotin labeled protein detected by Cy3 conjugated streptavidin Protein-Nucleic acid interactions Cy3 labeled genomic DNA Cy3 labeled mRNA Protein-Lipid interactions Biotin-conjugated liposome-phopshotidyl phosphate detected by Cy3 conjugated streptavidin * PI(3,4,5)P3 PC Calmodulin Genomic DNA Detection of different interactions on yeast proteome chips 蛋白质的分子结构可划分为四级，以描述其不同的方面： 一级结构：组成蛋白质多肽链的线性氨基酸序列。 二级结构：依靠不同氨基酸之间的C=O和N-H基团间的氢键形成的稳定结构，主要为α螺旋和β折叠。 三级结构：通过多个二级