第三四章细胞内蛋白质折叠新.pptxVIP

第三章:蛋白质折叠 第四章: 细胞内的蛋白质折叠;第三章:蛋白质折叠;二、关于蛋白质折叠的理论模型;三、蛋白质的去折叠(unfolding)和重折叠(refolding);加热, 酸或碱　盐, 破坏不牢固的次级键，使蛋白质功能丧失;核糖核酸酶;变性因素;蛋白质变性后：;复性;四、蛋白质折叠的热力学;五、蛋白质折叠的识别;六、蛋白质折叠识别的方式;另一种折叠的识别方式即所谓的Threading 。此方式主要是通过分析已知蛋白的由实验所测得的三维结构，来估计可允许的sub-in-del 而非进行未知结构与其同源家族的多重对齐。;第四章: 细胞内的蛋白质折叠;离体实验并不能精确地反映细胞内新生蛋白质的实际折叠过程.离体情况下蛋白质的去折叠后的再折叠(refolding)过程对蛋白质浓度及物理化学条件的要求完全不同于在细胞内所发生的情况 在离体情况下的再折叠实验往往涉及到整条肽链, 而在活体情况下则是当新生肽段的N 端在核糖体上一经合成或 刚从细胞膜上移位, 折叠活动就开始了。 3） 在细胞中许多蛋白质是以均一或不均一的寡体复合物就开始装配，而在离体的情况下当浓度如细胞内那么低时形成复合物的可能性极小;在细胞内至少存在两类蛋白质涉及到蛋白质的折叠这些蛋白被称作辅助蛋白(accessory protein);一） 二硫键异构酶(PDIase);一） 二硫键异构酶(PDIase) 结构;The enzyme PDI is a multi-domain, multi-functional member of the thioredoxin (a family of small redox active protein) superfamily. PDI can catalyse thiol-disulphide oxidation, reduction and isomerization. Isomerization occurs directly through intramolecular disulphide rearrangement or through cycles of reduction and oxidation.;Human PDI family;Yeast PDI PDB: 2B5E;Structure Function ;There are several hydrophobic patches of surrounding the active sites in the a and a’ domains and each of the β and β’ domains have one hydrophobic patch at the same relative sites. The primary substrate binding site has been mapped to the β’ domain for mammalian PDI. The hydrophobic patch on the β domain extends the substrate binding site of the β’ domain to form a larger substrate binding platform, which is located between the two active sites in the a and a’ domains.;Despite the high overall structure similarity, a few differences between the active sites are identified, which are likely to reflect the different catalytic properties of the a and a’ domain. The most noteworthy difference between the a and a’ domains is the redox state of their active site cysteines. The a domain is more stable in the oxidized form, while the a’ domain prefers to be in the reduced form. The equilibrium constant (Kox) for the oxidation of the cysteine pair in each active site by oxidized glutathione (GS