PROTEOMIKA. Rūta Navakauskienė. El.paštas:
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1 PROTEOMIKA Rūta Navakauskienė El.paštas:
2 Literatūra Simpson, Richard J. Proteins and proteomics: a laboratory manual. Cold Spring Harbor (N.Y.): Cold Spring Harbor. Laboratory Press, Walsh G. Proteins: Biochemistry and Biotechnology. New York: John Wiley & Sons, LTD, 2002 D. C. Liebler. Introduction to proteomics. Humana press R. E. March. An Introduction to Quadrupole Ion Trap Mass Spectrometry. J. Wiley & Sons Westermeier R., Naven T. Proteomics in Practice. Wiley-VCH Verlag GmbH, 2002.
3 1. Įvadas į Proteomiką 2. Proteomika genomikos dalis 3. Proteomikos Tipai: Proteomikos kurso struktūra (1) 3.1. Struktūrinė Proteomika Baltymų struktūra 3.2. Raiškos Proteomika Baltymų raiška 3.3. Funkcinė Proteomika Baltymų funkcijos - domenai ir modifikacijos
4 Proteomikos kurso struktūra (2) 4. Proteomo Nustatymo Metodologijos 4.1. Baltymų išskyrimas iš ląstelių kultūrų ir audinių bei jų frakcionavimas 4.2. Organelių proteomika 4.3. Baltymų skirstymo sistemos Chromatografija Baltymų elektroforezinis skirstymas SDS/PAGE Izoelektrofokusavimas Dvimatė elektroforezė Kapiliarinė elektroforezė DIGE sistema 4.4. Baltymų vaizdinimas ir skaitymas
5 Proteomikos kurso struktūra (3) 4.5. Masių spektrometrija Baltymų paruošimas masių spekrometrijai MS metodologijos ir techninė įranga:. MALDI TOF ESI MS/MS Baltymų nustatymas Duomenų analizė 4.6. Skaitiniai metodai proteomikoje 4.7. Proteomo tyrimų taikymas Ląstelių signaliniams keliams išaiškinti Medicininei diagnostikai ir prognostikai Vaistams kurti
6 ĮVADAS Į PROTEOMIKĄ
7 1994 m. Markas Vilkinsas (Marc Wilkins) pasiūlė terminą Proteomas tam tikro organizmo (ląstelės) sintetinamų baltymų visuma. Tai visų genomo koduojamų baltymų struktūros ir funkcijos nustatymas Tai viso organizmo baltymų rinkinio analizė. Jos metu nustatoma baltymų raiška jos kitimas organizmui vystantis ir sąveikaujant su aplinka. Proteomika tiria savitus ląstelės arba organizmo genų raiškos kitimus baltymų lygmenyje. Atliekama ne tik baltymų kokybinė ir kiekybinė analizė. Nustatoma ir jų vieta ląstelėje, modifikacijos, sąveikos bei funkcijos.
8 Kas yra proteomika? Proteomika tyrimų sritis, įgalinanti identifikuoti ir charakterizuoti visus baltymus, produkuojamus ląstelėje, audinyje ar organizme naudojant didelio našumo technologijas.
9 Proteomikos Tipai Struktūrinė Proteomika Baltymų struktūra rentgeno spindulių kristalografija NMR spektroskopija Raiškos Proteomika Baltymų sekvenavimas ir jų identifikavimas Elektroforezė, baltymų gardelės Masių spektrometrija Funkcinė Proteomika Baltymų modifikacijos Baltymų funkcijos nustatymas
10 Kodėl Proteomika? Žmogaus DNR nustatyta apie 25,000 genų, potencialiai koduojančių 25,000 skirtingų baltymų. Tačiau: alternatyvus RNR sukirpimas ir potransliacinės modifikacijos gali padidinti baltymų ar jų fragmentų skaičių iki 5 milijonų to pasekoje 1 genas gali koduoti apie skirtingų baltymų. Genominė informacija labai dažnai nesuteikia tikslios informacijos apie baltymų kiekį, struktūrą ar jo aktyvumą.
11 Molekulių Funkcijos Ląstelėje Mažos molekulės (pvz., amino rūgštys, vitaminai) ir jonai (pvz., chlorido, kalcio jonai): Perneša energiją (ATP) ir signalą (hormonai), Mažų molekulių vieta ir koncentracija ląstelėje yra kontroliuojama baltymų, esančių ląstelinėje membranoje (membraninės pompos, transporteriai ir jonų kanalai dalyvauja mažų molekulių ir jonų pernešime iš/į ląstelę ir jos organeles) Makromolekulės (angliavandeniai, baltymai ir nukleino rūgštys).
12 Makromolekulės Baltymai Suteikia Ląstelėms Struktūrą ir yra Atsakingi už Ląstelių Funkcijas Struktūriniai baltymai: citoskeleto (aktinas, tubulinas) chromatino (histonai) Signaliniai baltymai: citoplazminiai (Ras, MAPK) branduoliniai (AP-1, Jun, Fos) Membraniniai baltymai: receptoriai (EGF, Fas receptorius) transmembraniniai baltymai (dystroglikano kompleksas)
13 DNR mrnr Baltymas Metabolitas Genomika Transkriptomika Proteomika Metabolomika
14 Proteomikos tikslai Proteomika baltymų identifikavimo, jų struktūros bei galimų funkcijų apibūdinimo mokslas Baltymų identifikavimas, jų struktūrų suvokimas bei tarpusavio sąveikų nustatymas leistų: Pilnai išaiškinti proteomą; Prognozuoti, diagnozuoti bei išgydyti daug sunkių susirgimų; Geriau suprasti atskirų gyvų būtybių evoliucijos raidą.
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