第九讲 圆二色光谱及其应用.ppt

Instrumentation: synchrotron–based Synchrotron - whiz electrons around a ring. Can be used to produce very intense radiation by wiggling beam. Commonly used to produce X–rays (l around 0.1nm) But can be used to push signals down to 160nm and below Great for fast stopped-flow to see rapid changes Applications of CD in structural biology Determination of secondary structure of proteins that cannot be crystallised Investigation of the effect of e.g. drug binding on protein secondary structure Dynamic processes, e.g. protein folding Studies of the effects of environment on protein structure Secondary structure and super-secondary structure of membrane proteins Study of ligand-induced conformational changes Carbohydrate conformation Investigations of protein-protein and protein-nucleic acid interactions Fold recognition CD application Secondary structure of macromolecule a-helix a-helix: 19x nm (+) 208nm, 222nm (-) [q] x E-3 0 200 210 220 230 240 nm b-sheet: 195nm (+) antiparallel: red shift parallel: blue shift 215-7nm (-) b-sheet Unordered structure: (-) below 200nm (+) 218nm weak band unordered b-turn: 180-190nm (-) 200-205nm (+) 225nm (+) band, weak red shift 科学研究与开发 中科院某研究所根据HIV-1gp41上的两段七肽重复（heptad repeat，HR）序列1和2（HR1和HR2）对HIV-1的侵入有抑制作用，进行了HR1和HR2的多螺旋蛋白结构与抗病毒活性的研究，所设计的五螺旋蛋白的基因为HR12121。通过氨基酸连接子（Linker）将三个HR1的基因和二个HR2的基因连接在一起形成一种五螺旋蛋白HR12121的基因。圆二色谱分析证明HR12121和HR212蛋白发现这两种蛋白富含α螺旋结构，这表明HR12121和HR212蛋白非常稳定，不像多肽那样容易降解，并对HIV-1侵染细胞有很高的抑制活性。 目前我们已申请专利“一种抑制囊膜病毒感染的多螺旋蛋白及其编码基因与应用”。囊膜病毒含有许多引起人类重大疾病的病毒，其中抗艾滋病毒的蛋白药物是最重要的。 合作方式：有愿投资者，投资方可获得专利的独家使用权，约需1000-2000万元投资。签订意向书后，具体事宜面议。 克山病探究-硒缺乏 模型动物心肌线粒体膜的CD谱 [?] ? 正常对照 病区粮 病区粮,补加蔬菜 烟草花叶病毒侵染膜蛋白 引自:吉尚戎等 生物物理学报,2004,20 Unfolding/folding 216nm 222nm Conformational varies 190 200 210 220 230 240 250 260 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 2.0 a5 in 5 % SDS/PBS a5 in 50 % TFE/PBS a5 in pH7 a5 in 50 % TFE/H 2 O a5 in H 2 O [|è]X1E-4 deg.cm 2 /dmol -1 wavelength(nm) 216nm   CD spectra of GCN4 leucine zip