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Zakaj smo tu?

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Zakaj smo tu? OECD Biotechnology Statistics 2009 (http://www.oecd.org/sti/42833898.pdf)
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Page 1: Zakaj smo tu?

Zakaj smo tu?

OECD Biotechnology Statistics 2009 (http://www.oecd.org/sti/42833898.pdf)

Page 2: Zakaj smo tu?

Proteomika

“Kot da geni in mRNA kaj pomenijo...”

Page 3: Zakaj smo tu?

Nature (2006) 441:840

DNA microarray

Single-cell proteomics (flow cytometry)

PROTEOMIKA

Page 4: Zakaj smo tu?

“I think the problem, to be quite honest with you, is that you’ve never actually known what the question is.”

Life?

Universe?

Everything?

Douglas N. Adams

1952-2001

Page 5: Zakaj smo tu?

Eksperimenti brez a priori hipoteze

Fishing expeditions vs. Postavljanje pravih vprašanj

Page 6: Zakaj smo tu?
Page 7: Zakaj smo tu?

http://www.ibgc.u-bordeaux2.fr/YPM

15kD

180kD

pI M

w

6,8 4,2

Page 8: Zakaj smo tu?

Merjenje razlik v izražanju proteinov: ‘Difference Gel Electrophoresis (DiGE)’

2-D Electrophoresis. Principles and Methods; GE Healthcare

Page 9: Zakaj smo tu?

Masna spektromerija v proteomiki

Nature (2003) 422:198

Page 10: Zakaj smo tu?

Masna spektromerija v proteomiki

Nature (2003) 422:198

Page 11: Zakaj smo tu?

Nature (2013) 494:266

Proteom kot kvantitivna lastnost (fenotip)

Page 12: Zakaj smo tu?

Post-translacijske modifikacije proteinov

Molecular Cell (2008) 31:449

Page 13: Zakaj smo tu?

A plastic epigenome. DNA and histones are targets of multiple modifications that convey

flexibility to the genome.

P Sassone-Corsi Science 2013;339:148-150

Published by AAAS

Page 14: Zakaj smo tu?

Nature Reviews Mol Cell Biol (2013) 14:211

Page 15: Zakaj smo tu?

Primer analize ‘fosfoproteoma’

Page 16: Zakaj smo tu?

Molecular Systems Biology (2012) 8:623

Science (1997) 278:680

Page 17: Zakaj smo tu?

Acetilacija proteinov / ‘acetilom’

Molecular Cell (2013) 49:1

Page 18: Zakaj smo tu?

Interakcije protein-DNA (ChIP-chip, ChIP-seq)

Page 19: Zakaj smo tu?

ENCODE - NGS in proteomika

Molecular Systems Biology (2013) 9:640

Page 20: Zakaj smo tu?

Cell (2013) 152:642

Page 21: Zakaj smo tu?

Določanje prisotnosti/količine : določanje funkcije

- imunoprecipitacija s pomočjo TAP označevalca/MS (proteinski kompleksi)

- dvohibridni sistem kvasovke (Y2H, protein-protein fizične interakcije)

- proteinske mikromreže

Nature (2003) 422:208

Page 22: Zakaj smo tu?

Test s komplementacijo fragmentov proteina (Protein-fragment complementation assay; PCA)

Membranski kvasni dvohibridni sistem

FRET/BRET ( Fluorescence/bioluminescence resonance-energy transfer )

Curr Opinion Biotech (2011) 22:50

Kvasni dvohibridni sistem

Page 23: Zakaj smo tu?

Nature (2003) 422:208

Strategije Y2H

Page 24: Zakaj smo tu?

Nature Methods (2011) 8:478

Y2H & NGS

Page 25: Zakaj smo tu?

Nature (2002) 415:141

Analiza proteinskih kompleksov z afinitetno kromatografijo / MS

Page 26: Zakaj smo tu?
Page 27: Zakaj smo tu?

Primer: Piruvat kinaza – lokalizacija in acetilacija

Mol Cell (20013) 52:340

Page 28: Zakaj smo tu?

J Proteom Res (2009) 8:20

High content microscopy

Page 29: Zakaj smo tu?

Pex11 represents an ancestral module from which multidomain nuclear receptors arose

TiBS (2000) 25:227

Nature Rev Mol Cell Biol (2010) 11:644

Pex11 is required for peroxisome proliferation

Fis1

Pex11

- the most abundant protein in the px membrane

- deletion prevents and over-expression induces px proliferation

- recruits, but does not necessarily binds to, (mt) fission machinery proteins

Page 30: Zakaj smo tu?

MYTH Nature Protocols (2010) 5:1281-1293

How is the signal (from Pex11) transmitted?

cy

N-

C-

Pex11 Ftp1 (Pex34)

N-

C-

Fis1

N-

C-


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