1、Protein foldingJames Dewey WasonFrancis Harry Compton Crick DNA RNA Proein?中國啟動人類肝臟蛋白質(zhì)組計劃國際人類蛋白質(zhì)組計劃的以上的任務(wù)。proteomics為什么要開展蛋白質(zhì)折疊的研究?研究的源動力 disease 免疫病毒Protein folding & live蛋白質(zhì)研究先驅(qū)吳 憲 1893,11,24-1959,8,8年到美國麻省理工學(xué)院。因憤于我國海軍落后，初學(xué)造船工程。因受赫胥黎“生命的物理基礎(chǔ)”一文的影響，年后改習(xí)化學(xué)。年畢業(yè)，獲理學(xué)學(xué)士學(xué)位，留校任化學(xué)系助教。年進哈佛大學(xué)醫(yī)學(xué)院生物系，成為美國著名生物化

2、學(xué)家 Folin教授的研究生，進行血液化學(xué)研究。1924年起用各種方法使蛋白質(zhì)變性，1931年得出如下理論：蛋白質(zhì)的變性是由于蛋白質(zhì)分子由折疊而變?yōu)槭嬲埂?變性劑巰基乙醇復(fù)性ribonuclease Some denatured proteins can be renatured denatured moleculeAnfinsen原理 1961Probability that correct folding would occur in ribonuclease given that there are 8 cysteine residues 1/7 x 1/5 x 1/3 x 1 = 1/

3、105expected activity 1%observed activity was 100%if one conformation is explored every 0.1 psec, then time to refold (t) = 1087 sec2n torsion angles can have 32n 10n possible conformationsdirected pathways of folding must existsif n =100, then number of conformations, 10100 Mechanisms to explain re-

4、folding Factors driving protein foldingUnfolded stateFormation of elements of 2-stru.Framework modelFolded con.Assembly of 2-stru. gabcdefgabcdefgabcdefgabcdefgabcdSTHMKQLEDKVEELLSKNYHLENEVARLKKLVGERGCN4 leucine zipper CD spectra of GCN4 leucine zipper in the presence of different concentrations of

5、SDSSDS4 M GuHClChanges of ellipticity at 222 nm in the presence of different concentrations of SDSNative gel electrophoresis of leucine zipper treated with SDS of different concentrationsLane 1 was the native leucine zipper peptide (control); lanes 2- 6 were samples treated with 0.1, 0.2, 0.3, 0.6,

6、and 1.0 mM SDSsome lead straight downhillEnergy surfaces to visualize protein folding pathwaysAa more realistic energy landscapethe protein is funneled towards a native statemany pathways are possibleothers may lead to energyminima that delay proper foldingChanges of fluorescence emission spectra of

7、 Tg denatured in various concentrations of GuHClThyroglobulinANS binding characteristics of Tg in various GuHCl concentrations蛋白質(zhì)功能區(qū)肌酸激酶活性部位熒光探針暴露的速度常數(shù)鹽酸胍 （M）熒光OPTA內(nèi)源熒光失活k1k2k1k20.30.380.0490.0150.51.180.110.00383.60.0031.02.90.044.3酶活性部位的柔性學(xué)說鄒承魯 Proteinprotein interface design erythropoietin EPO-EP

8、ORERPH1-EPORLiuS,LiuSY,ZhuXL, LiangHH,CaoAN,ChangZJandLaiLH*. Nonnaturalprotein-proteininteraction-pairdesignbykeyresiduesgrafting.PNAS,2007,104,5330 藥物研究 抗氧化劑對A1-40結(jié)構(gòu)的影響庾照學(xué)等,中國病理生理雜志,2000,16FT - IR spectra of A1 - 40 in PBS(pH7. 4) for 30minFT - IR spectra of A1 - 40 in PBS(pH7. 4) for 7 daysFT - I

9、R spectra of A1 - 40 in PBS(pH7. 4) for 7 daysFT- IR spectra of A1 - 40 in PBS(pH7. 4) with TA9901 for 7 days (percent ratio : A1 40 :TA9901 = 11)衰 減 全 反 射 紅 外 光 譜研究人乳腺癌組織A1635/A1652A1625/A1652A1645/A1652A1662/A1652A1682/A1652benign0.980.650.640.490.17malignant0.430.230.560.370.09開闊思路記憶合金棒矯正脊柱側(cè)凸 盧世璧

10、(院士)HSP 腫瘤疫苗分子伴侶molecular chaperones 新生肽鏈的折疊Misfolding & Protein Conformational Disorders瘋牛病帶給生命科學(xué)界的思考Mad cowTSEAlzheimers D. Amyloid Protein & Tau proteinFamilial visceral Amyloidosis Lysozyme.Parkinson D. -synuclein Huntington D. Glutamine-repeatPrion D. Prion protein Sickle cell anaemia Haemoglob

11、inConformational BrainsDisordersProteinConforma-tionalDisorders (PCD)Human Prion DiseasesSporadic formCreutzfeldt-Jakob disease (CJD)Familial (inherited) formFamilial CJDFatal familial insomniaGerstmann-Straussler-Scheinker syndromeAcquired (transmitted) formIatrogenic CJDKuruNew Variant CJD (relate

12、d to Mad Cow Disease)Animal Prion DiseasesScrapieSheep and goatBovine spongiform encephalopathy (Mad Cow Disease)CattleFeline spongiform encephalopathyCat (domestic cats, cheetahs, pumas)Transmissible mink encephalopathy MinkChronic wasting diseaseMule deer, elk朊病毒(Prion)病的共同特征 臨床表現(xiàn):癡呆、共劑失調(diào)、震顫等癥狀 病理

13、學(xué)上的特點:大腦皮層的神經(jīng)原細胞退化、空泡變性、死亡、消失，星狀膠質(zhì)細胞增生，蛋白酶抗性的PrP積聚，有時產(chǎn)生淀粉樣斑 Prion Protein Gene (PRNP)-Located on chromosome 20 in humans, chromosome 2 in mouse-Encodes a glycoprotein with two sites for N-linked oligosaccharites and a C-terminal GPI anchor-High expression in brain. Lower expression in peripheral tis

14、sues-10-15% of all cases are familial. About 20 mutations are linked to familial disease.The Nobel Prize in Physiology or Medicine 1976for their discoveries concerning new mechanisms for the origin and dissemination of infectious diseasesBaruch S. BlumbergD. Carleton GajdusekStanley B. PrusinerThe N

15、obel Prize in Physiology or Medicine 1997for his discovery of Prions - a new biological principle of infectionDNA RNA ProteintranscriptiontranslationSequence structure functionfolding二個中心法則1. Genetics:2. Protein:中心法則?Conformational transition: from alpha-helix rich to beta-sheet richPrPc PrPscPrPcPrPscPrP27-30ReferencesRoger H.Pain， Mechanisms of Protein