基因組和蛋白組學技術-蛋白質組學ppt課件

1、DNAmRNA蛋白質轉錄翻譯基因蛋白質細胞特異性基因表達生理狀態(tài)蛋白質相互作用溫度應激狀態(tài)培養(yǎng)條件藥物作用數(shù)量有限結構相對穩(wěn)定復雜性多變性直接反映生命現(xiàn)象復雜的調(diào)控1994年，澳大利亞Williams提出蛋白組學，研究不同時空狀態(tài)蛋白質整體變化。蛋蛋 白白 質質 組組6實驗室服務發(fā)表文章7、Jun Pang, Weipeng Liu, Xiaopeng Liu, Liaoyuan Li, Youqiang Fang, Qipeng Sun, Shaojun Liu, Mingtao Li, Zulan Su, Xin Gao. Profiling protein markers associa

2、ted with lymph node metastasis in prostate cancer by DIGE-based proteomics analysis. J Proteome Res. 2019;9:216-226.8、Yan-Xin Luo, Ji Cui, Lei Wang,Dian-Ke Chen. Identification of cancer-associated proteins byproteomics and downregulation of b-tropomyosinexpression in colorectal adenoma and cancer.

3、Proteomics Clin. Appl. 20219、Jing Luo, Jine Yang, Bing-Yun Yu, Wei Liu, Mingtao Li, Shi-Mei Zhuang. Identification of Siah-interacting protein as a potential regulator of apoptosis and curcumin resistance. Oncogene. 2019; 29(48):6357-636610、Liu W, Zhou XW, Liu S, Hu K, Wang C, He Q, Li M. Calpain-tr

4、uncated CRMP-3 and -4 contribute to potassium deprivation-induced apoptosis of cerebellar granule neurons. Proteomics. 2021 Jul;9(14):3712-28.11、Wang J, Wu S, Jin X, Li M, Chen S, Teeling JL, Perry VH, Gu J. Retinoic acid-inducible gene-I mediates late phase induction of TNF-alpha by lipopolysacchar

5、ide. J Immunol. 2019 Jun 15;180(12):8011-9.12、Kang L, Yang ZL, Liu W, Zhang LJ, Liu SJ, Huang MJ, Li MT, Wang JP. (2019) Serum proteomic variation study in patients with Crohn disease. Zhonghua Wei Chang Wai Ke Za Zhi. 2019 May;11(3):266-9. 13、Fu H, Dou J, Li W, Luo J, Li KC, Lam CS, Lee NT, Li M, H

6、an Y. Mecamylamine prevents neuronal apoptosis induced by glutamate and low potassium via differential anticholinergic-independent mechanisms. Neuropharmacology. 2019 Mar;54(4):755-65.14、Hongjun Fu, Wenming Li, Jialie Luo, Nelson T.K. Lee, Mingtao Li, Karl W.K. Tsim, Yuanping Pang, Moussa B.H. Youdi

7、m and Yifan Han. (2019) Promising anti-Alzheimers dimer bis(7)-tacrine reduces -amyloid generation by directly inhibiting BACE-1 activity. Biochem Biophys Res Commun. 2019 Feb 15;366(3):631-615、Ma C, Ying C, Yuan Z, Song B, Li D, Liu Y, Lai B, Li W, Chen R, Ching YP, Li M (2019) dp5/HRK is a c-Jun t

8、arget gene and required for apoptosis induced by potassium deprivation in cerebellar granule neurons. J Biol Chem 282:30901-3090916、Mei, Y, Yuan, Z, Song, B, Li, D, Ma, C, Hu, C, Ching,YP, Li, M . Activating Transcriptional Factor 3 up-regulated by JNK/c-Jun contributesto apoptosis induced by potass

9、ium deprivation in cerebellar granule neurons, Neuroscience.2019,17、Wang W, Yang Y, Ying C, Li W, Ruan H, Zhu X, You Y, Han Y, Chen R, Wang Y, Li M .(2019) Inhibition of glycogen synthase kinase-3beta protects dopaminergic neurons fromMPTP toxicity. Neuropharmacology 52:1678-168418、Yuan Z, Mei Y, Zh

10、ou J, Tan M, Song B, Ma C, Ying C, Li D, Ching YP, Li M . (2019)E2F1is not essential for apoptosis induced by potassium deprivation in cerebellar granule neurons.Neurosci Lett 424:155-159.19、Guo QC, Shen JN, Jin S, Wang J, Huang G, Zhang LJ, Huang G, Yin JQ, Zou CY, LiMT(2019) Comparative proteomic

11、analysis of human osteosarcoma and SV40-immortalizednormal osteoblastic cell lines. Acta Pharmacol Sin. 28:850-858.20、Li W, Pi R, Chan HH, Fu H, Lee NT, Tsang HW, Pu Y, Chang DC, Li C, Luo J, Xiong K, LiZ, Xue H, Carlier PR, Pang Y, Tsim KW, Li M, Han Y*(2019) Novel dimeric AChE inhibitorBis(7)-tacr

12、ine, but not Donepezil, prevents glutamate-induced neuronal21、Li C, Carlier PR, Ren H, Kan KK, Hui K, Wang H, Li W, Li Z, Xiong K, ClementEC, Xue H, Liu X, Li M, Pang Y, Han Y* (2019) Alkylene tether-length dependent g-aminobutyric acid type A receptor competitive antagonism by tacrine dimmers.Neuro

13、pharmacology. 52:436-443.22、Li W, Xue J, Niu C, Fu H, Lam CS, Luo J, Chan HH, Xue H, Kan KK, Lee NT, Li C,Pang Y, Li M, Tsim KW, Jiang H, Chen K, Li X, Han Y* (2019) SynergisticNeuroprotection by Bis(7)-tacrine via Concurrent Blockade of N-Methyl-D-aspartateReceptors and Neuronal Nitric Oxide Syntha

14、se. Mol Pharmacol 71:1258-1267.23、Xi L, Xu X, Liu W, Li X, Liu Y, Li M, Zhang J, Li M(2019) Differentially expressedproteins of pathogenic Penicillium marneffei in yeast and mycelial phases. J MedMicrobiol 56:298-304. 24、Zhang AX, Yu WH, Ma BF, Yu XB, Mao FF, Liu W, Zhang JQ, Zhang XM, Li SN,Li MT,

15、Lahn BT, Xiang AP*(2019)Proteomic identification of differently expressedproteins responsible for osteoblast differentiation from human mesenchymal stem cells.Mol Cell Biochem. 304:167-17925、Song B, Ma C, Gong S, Yuan Z, Li D, Liu W, Li W, Chen R, Zhu X, Zeng J, Han Y,Li M (2019) Extracellular signa

16、l-regulated kinases are not involved in activity dependentsurvival or apoptosis in cerebellar granule neurons. Neurosci Lett 407: 214-218.26、Yang LL, Liu XQ, Liu W, Cheng B*, Li M . (2019)Comparative analysis of wholesaliva proteomes for the screening of biomarkers for oral lichen planus. Inflamm Re

17、s55:405-407.27、Li W, Lee NT, Fu H, Kan KK, Pang Y, Li M, Tsim KW, Li X, Han Y*(2019)Neuroprotection via inhibition of nitric oxide synthase by bis(7)-tacrine. Neuroreport17:471-474.28、Fu H, Li W, Lao Y, Luo J, Lee NT, Kan KK, Tsang HW, Tsim KW, Pang Y, Li Z,Chang DC, Li M, Han Y*(2019) Bis(7)-tacrin

18、e attenuates beta amyloid-induced neuronalapoptosis by regulating L-type calcium channels. J Neurochem 98:1400-1410.29、Yang M, Liu W, Wang CY, Liu T, Zhou F, Tao J, Wang Y*, Li M(2019) Proteomicanalysis of differential protein expression in early process of pancreatic regeneration inpancreatectomize

19、d rats. Acta Pharmacol Sin 27:568-578.30、Deng R, Li W, Guan Z, Zhou JM, Wang Y, Mei YP, Li M, Feng GK, Huang W, Liu ZC, Han Y, Zeng YX, Zhu XF*(2019) Acetylcholinesterase expression mediated byc-Jun-NH2-terminal kinase pathway during anticancer drug-induced apoptosis. Oncogene25:7070-7077.31、Li J*,

20、Tan Z, Li MT, Liu YL, Liu Q, Gu XF, Zhou JZ, Zhuang GL.(2019) Study ofaltered expression of annexin IV and human endometrial receptivity Zhonghua Fu ChanKe Za Zhi. 41:803-5. Chinese.32、Shi L, Gong S, Yuan Z, Ma C, Liu Y, Wang C, Li W, Pi R, Huang S, Chen R, Han Y,Mao Z, Li M . (2019) Activity depriv

21、ation-dependent induction of the proapoptotic BH3only protein Bim is independent of JNK/c-Jun activation during apoptosis in cerebellargranule neurons. Neurosci Lett 375:7-12. 33、. Wang X, Tang X, Li M, Marshall J, Mao Z*(2019) Regulation of neuroprotectiveactivity of myocyte enhancer factor 2 by cA

22、MP-PKA signaling pathway in neuronalsurvival. J Biol Chem 280:16705-167013. 34、 Wiedmann M, Wang X, Tang X, Han M, Li M, Mao Z*(2019) PI3K/Akt-dependentregulation of the transcription factor myocyte enhancer factor-2 in insulin-like growthfactor-1- and membrane depolarization-mediated survival of ce

23、rebellar granule neurons. JNeurosci Res 81:226-234.35、Wang W, Shi L, Xie Y, Ma C, Li W, Su X, Huang S, Chen R, Zhu Z, Mao Z, Han Y, Li M . (2019). SP600125, a new JNK inhibitor, protects dopaminergic neurons in the MPTP model of Parkinsons disease. Neurosci Res. 48:195-202.36、Wang W, Ma C, Mao Z, Li

24、 M . (2019) JNK inhibition as a potential strategy in treating Parkinsonss disease. Drug News Perspect. 17:646-654. 28. Xie Y, Liu Y, Ma C, 37、Yuan Z, Wang W, Zhu Z, Gao G, Liu X, Yuan H, Chen R, Huang S, Wang X, Zhu X, Wang X, Mao Z, Li M . Indirubin-3-oxime inhibits c-Jun NH2-terminal kinase: anti

25、-apoptotic effect in cerebellar granule neurons. Neurosci Lett. 2019; 367(3):355-359. 37、Pi R, Li W, Lee NT, Chan HH, Pu Y, Chan LN, Sucher NJ, Chang DC, Li M, Han Y*(2019) Minocycline prevents glutamate-induced apoptosis of cerebellar granule neurons by differential regulation of p38 and Akt pathwa

26、ys. J Neurochem 91:1219-1230.38、. Yun JP*, Liew CT, Chew EC, Yin XY, Lai PB, Fai YH, Li HK, Jin ML, Ding MX, Li MT, Lin HL, Lau WY(2019) Nuclear matrix protein expressions in hepatocytes of normal and cirrhotic rat livers under normal and regenerating conditions. J Cell Biochem. 91:1269-1279.9、張劍文，劉

27、 煒，傅斌生，胡坤華，劉少軍，李華，張琪，黎明濤，陳規(guī)劃 乙腈預處理血清尋找肝細胞癌發(fā)病相關的蛋白質 中山大學學報 醫(yī)學科學版 第31卷 第2期 2019 年 3月10、傅斌生，劉煒，張劍文，張彤，李華，陳規(guī)劃. 肝癌轉移相關分子的血清蛋白質組學分析. 南方醫(yī)科大學學報, 2021, 29(9):1775-1778.胡志成，朱家源，朱 斌，郭 棟，陳 斌，張 凱，胡坤華，黎明濤，唐冰 病理性瘢痕與正常皮膚的蛋白質組學研究 中國病理生理雜志 2019，27( 9) : 18021806。11、吳曉濱，彭俊生，李初俊，王輝，杜艷平，楊祖立，向軍，胡坤華，劉煒, 黎明濤 蛋白質組學檢測結直腸腺瘤及早

28、期惡變患者血清的差異蛋白質變化及意義 中國病理生理雜志2009 , 25( 6): 1098- 1103。12、梁瓊，李揚，王連唐，劉郁林，羅燦橋，梁惠珍，黎明濤 骨肉瘤組織與良性骨腫瘤的比較蛋白質組學分析 中國病理生理雜志2009 , 25( 6): 1089- 1093。13、朱永華，劉煒，石忠松，劉少軍，李明昌，黎明濤，潘偉生，黃正松 顱內(nèi)動脈瘤患者血清蛋白組變化的研究 中山大學學報(醫(yī)學科學版) 第27卷 第6期2019 年 11 月14、崔冀，汪建平，彭俊生，黃奕華，何裕隆，蘭平，蔡世榮，駱衍新，胡坤華，劉煒，黎明濤 結腸癌的蛋白質組學研究 中國病理生理雜志 2019 , 24( 5

29、): 1013- 1017Sample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab Workflow1、蛋白提取組織、血清、細胞等）2、蛋白純化丙酮沉淀法）3、蛋白定量等量分析）Sample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Han

30、ding WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab Workflow熒光標記技術2-D DIGE）雙向電泳分離比較類別類別樣本數(shù)樣本數(shù)染色時間染色時間可控性可控性內(nèi)標內(nèi)標靈敏度靈敏度熒光3分離前參數(shù)，特定波長有最高考、銀染1分離后目測，經(jīng)驗無-Cy2(內(nèi)參內(nèi)參)Cy3Cy5內(nèi)標cy2）樣品1cy3）樣品2cy5）內(nèi)標cy2）樣品3cy3）樣品4cy5）內(nèi)標使用，數(shù)據(jù)更精確Sample preparation2-D electrophoresis/DIGEdetection & i

31、mage analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab WorkflowSample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab Workflow考染考染銀染銀染熒光熒光高靈敏高靈敏兼兼 容容

32、高通量高通量/高效率高效率Sample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab Workflow1. 二級分析，蛋白鑒定率高2. 蛋白質、多肽等分子量精確測定質譜鑒定1233. 6851631. 8771781. 0191965. 0502729. 4651441. 6982224. 073831. 4781118. 5641100

33、. 861050010001500Intens. a.u.100015002000250030003500m/z1233蛋白斑點肽段1級質譜：肽段質量2級質譜：氨基酸序列數(shù)據(jù)庫比對，鑒定酶解：特異性切割L,R1233ELLSQYDFPGDDTPIVRELLSQYDFPGDDTPIVRELLSQYDFPGDDTPIVELLSQYDFPGDDTPI123311911077Sample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinf

34、ormaticsProtein function studyProteomics Lab Workflow40414243444546Sample preparation2-D electrophoresis/DIGEdetection & image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab Workflow功 能 研 究52Sample preparation2-D electrophoresis/DIGEdetection &

35、; image analysisSpot Handing WorkstationMALDI-ToF/ToFBioinformaticsProtein function studyProteomics Lab WorkflowCalpain鈣蛋白酶底物調(diào)控凋亡的蛋白質凋亡刺激凋亡刺激細胞凋亡實例1蛋白被calpain剪切的模式圖Gel no: 67913Standard25k 5k Z5k StandardZ5k5kStandard5kZ5kStandardZ5k25kStandard25k5kStandard25kZ5kStandard5k25kStandard25kZ5kStandard5k

36、25kStandardZ5k5kStandardized Log Abundance50.10.050-0.05-0.1-0.15-0.2-0.25Gel no: 72081Standard25k 5k Z5k StandardZ5k5kStandard5kZ5kStandardZ5k25kStandard25k5kStandard25kZ5kStandard5k25kStandard25kZ5kStandard5k25kStandardZ5k5kStandardized Log Abundance0-0.05-0.1-0.15-0.2-0.25-0.3-0.35-0.4-

37、0.45-0.5-0.55-0.6蛋白原型切割產(chǎn)物cy2cy3cy5Ubiquitin carboxyl-terminal hydrolase isozyme L1 蛋蛋白質降解白質降解Proteasome subunit alpha type 5 蛋白蛋白質降解質降解被剪切的蛋白質及其片段Name MW pI CRMP-1 62.5 6.6CRMP-2 64 6.1CRMP-4 62 5.8MAP-1B 270 4.7 MW pI 57 5.9 58 5.8 58 6.0 63 5.6CRMP- 4 被剪切Gel no: 67913Standard25k 5k Z5k StandardZ5k

38、5kStandard5kZ5kStandardZ5k25kStandard25k5kStandard25kZ5kStandard5k25kStandard25kZ5kStandard5k25kStandardZ5k5kStandardized Log Abundance50.10.050-0.05-0.1-0.15-0.2-0.25Gel no: 72081Standard25k 5k Z5k StandardZ5k5kStandard5kZ5kStandardZ5k25kStandard25k5kStandard25kZ5kStandard5k25kStandard25k

39、Z5kStandard5k25kStandardZ5k5kStandardized Log Abundance0-0.05-0.1-0.15-0.2-0.25-0.3-0.35-0.4-0.45-0.5-0.55-0.6Western blot 驗證25 K 5 K 5 KALLM CRMP-1CRMP-2CRMP-3CRMP-42021, Proteomics69實例2局限型：預后良好轉移性：惡性，預后差缺乏明確診斷喪失治療時機，浪費人力物力70Cy2(內(nèi)參內(nèi)參)Cy3(局限型局限型)Cy5(轉移型轉移型)實例471局限型局限型轉移型轉移型CRMP-4 72CRMP-4 在轉移型中下調(diào)局限型

40、局限型轉移型轉移型L6J Proteome Res 2019 9 216-262019, J Proteome Res74 Oncogene 2019 29 4555-66 前列腺癌治療靶標前列腺癌治療靶標 2019, Oncogene761 1、20192019年廣東省科技進步一等獎年廣東省科技進步一等獎 前列腺癌的分子診斷前列腺癌的分子診斷2 2、20192019年國家科技進步獎一等獎年國家科技進步獎一等獎前列腺癌診療體系的創(chuàng)新及其關鍵技術的應用前列腺癌診療體系的創(chuàng)新及其關鍵技術的應用3 3、專利、試劑盒、專利、試劑盒( (已測九百例研發(fā)和產(chǎn)業(yè)化已測九百例研發(fā)和產(chǎn)業(yè)化用于前列腺癌轉移早期診

41、斷的試劑盒用于前列腺癌轉移早期診斷的試劑盒專利公告號：CN102051408A 77馬內(nèi)菲青霉菌致病的蛋白質組學研究實例378Cy2(內(nèi)參內(nèi)參)Cy3(酵母相酵母相)Cy5(菌絲相菌絲相)紅色Cy5）：菌絲相綠色Cy3）：酵母相2019, J Med Microbiol實例4淀粉樣血清相關蛋白SAA2019, Proteomics Clin Appl1、差異模型差異窗口盡可能大） 病理形態(tài)學改變 臨床診斷 生物學指標2、意義：設計合理樣本數(shù)據(jù)采集樣本數(shù)據(jù)采集4212.55339.41946.52673.23886.14646.59292.07768.15906.26633.5010002000300040005000Intens. a.u.0200040006000800010000m/z分組分組例數(shù)例數(shù)正常組50疾病組50建模方法建模方法模型識別模型識別率（率（%）交叉驗證交叉驗證能力能力(%)基因遺傳算法98.4591.83快速分類算法91.2893.6835例樣本驗證，診斷率87%0250500750100012501500Inte