Presentasi sedang didownload. Silahkan tunggu

Presentasi sedang didownload. Silahkan tunggu

Proteomik. Proteomics Proteomics (PROTEOME Analysis) – analysis of the entire PROTEin complement expressed by a genOME (Wilkins et al., 1996). Proteomics.

Presentasi serupa


Presentasi berjudul: "Proteomik. Proteomics Proteomics (PROTEOME Analysis) – analysis of the entire PROTEin complement expressed by a genOME (Wilkins et al., 1996). Proteomics."— Transcript presentasi:

1 Proteomik

2 Proteomics Proteomics (PROTEOME Analysis) – analysis of the entire PROTEin complement expressed by a genOME (Wilkins et al., 1996). Proteomics (PROTEOME Analysis) – analysis of the entire PROTEin complement expressed by a genOME (Wilkins et al., 1996). Professor Maxey C.M. Chung

3 TIME Magazine July 3, 2000

4 Ctr 12 h 24 h 72 h Hsp 27 Cathepsin D Colorectal cancer cell lines treated with Butyrate, a HDAC inhibitor Traditional Western Blot (single candidate analysis) A)B) Proteomics refers to the large-scale study of the structure & functions of proteins. Proteomics A) Normal Liver B) Hepatocellular carcinoma

5 Proteomics versus traditional approach

6 The Proteome: A snapshot of proteins in the cell Normal Lesion In Situ Invasive Cancer Metastatic Cancer

7 Why Proteomics? 1.The genome is a rather constant entity whereas the proteome is dynamic. (differs from cell to cell, is constantly changing through its interactions with the genome and the environment) 2.Proteins, not genes, are the functional workhorses of a cell. 3.In addition, there is a poor correlation between mRNA levels and protein levels in the cell. 4.Proteomics also allows for the identification of post-translational modifications (eg phosphorylations) which directly regulate activity by switching on/off proteins. 5.Biofluids (eg blood, GI fluids, urine, CSF, amniotic fluid) do not have corresponding genomes or transcriptomes from which gene expression can be analysed.

8 NUMBER OF PROTEINS IN A PROTEOME 1.20,000 – 40,000 genes in human Each gene produces 3-6 final modified proteins Post-translational modifications (PTMs) Alternate splicing 2.Number of proteins in higher organism is therefore between 60,000 to 250, A single genome can give rise to different proteomes depending on such variables such as cell cycle, stress response, pathological conditions

9

10 PROTEOMICS The proteome is a dynamic entity Protein changes resulting from drug treatment, diseases etc This could be a result of differences in expression and/or post-translational modifications Note that the GENOME is static

11 Functional Validation Clinical Validation Pipeline for Clinical Proteomics Sample Collection Biomarkers / Drug targets / Mechanisms Sample Preparation Protein Profiling and separation Protein Identification by Mass Spectrometry Bioinformatics

12 Protein Elektroforesis Gel Poliakrilamid SDS-PAGE Pergerakan molekul bermuatan dalam medan listrik Sodium lauryl sulfate membentuk kompleks bermuatan negatif dengan protein 2-merkaptoetanol untuk memutus ikatan disulfida Pemisahan berdasarkan massa molekul

13 Elektroforesis 2-D: Pemisahan protein ( jenis) berdasarkan muatan dan ukuran

14 Western blot: identifikasi protein menggunakan antibodi yang spesifik

15

16 Proteomics Jun;6(11): Clinical Proteomics in O&G Spots differentially expressed in patients with RSA Detection of candidate biomarkers in follicular fluid of controls and RSA patients by western blots Fibrinogen chains,  and  Antithrombin

17 HPLC: Pemisahan Campuran Protein

18 Protease: hidrolisis ikatan peptida

19

20 Spektrofotometri Massa: Penentuan massa molekul Pembentukan ion. Konversi molekul dalam bentuk padat atau larutan menjadi ion dalam fasa gas Pemisahan ion dalam fasa gas berdasarkan m/z. Detektor. Penentuan m/z setiap ion

21 Matrix-Assisted Desorption Ionization (MALDI) Protein dicampur dengan komponen matriks (Asam dihidroksi benzoat) Sinar 337 nmmengeksitasi matriks dan protein melalui mekanisme transfer energi dari matriks ke sampel protein MALDI menghasilkan ion [M+H] +

22 Electrospray Ionization (ESI) Tegangan listrik tinggi (3-5 kv) pada larutan Pengupan pelarut (dengan gas hidrogen yang dipanaskan) Menghasilkan ion bermuatan lebih dari satu [M+nH]+n

23 Surface enhaced laser desorption ionizaiton (SELDI). Profil protein dari cairan tubuh (darah)

24 Spektrofotometri Peptida

25 Tandem Mass Spectroscopy Peptida DNA polymerase

26 Spektrofotometri massa: Kuantitatif Peptida

27 Diagnosa Pola Proteom

28 Protein Tagging System: Isolasi dan Pemurnian Protein

29 Interaksi Protein-Protein Sistem Dua Hibrida Ragi

30

31

32 Interaksi Protein dengan Koimunopresipitasi


Download ppt "Proteomik. Proteomics Proteomics (PROTEOME Analysis) – analysis of the entire PROTEin complement expressed by a genOME (Wilkins et al., 1996). Proteomics."

Presentasi serupa


Iklan oleh Google