© Copyright Ebiointel,SL 2006 Proteómica Imma Ponte.

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Presentation transcript:

© Copyright Ebiointel,SL 2006 Proteómica Imma Ponte

© Copyright Ebiointel,SL 2006 Proteómica: Estudio de los proteomas Proteomas: Proteinas codificadas por un genoma Proteómica: Introducción 1 genoma diferentes proteomas genes/genoma Proteïnas/genoma Proteïnas/celula

© Copyright Ebiointel,SL 2006 Proteomas: Separacion, identificación y caraterización de las proteínas presentes en una muestra biologica Proteómica: Introducción Estado metabólico Momento del desarrollo Niveles de expresión Modificaciones postranscripcionales Localización celular Activación por señales externas

© Copyright Ebiointel,SL 2006 Ejemplos de Proteomas: Proteómica: Introducción

© Copyright Ebiointel,SL 2006 Comparar proteomas distintos Identificar una proteína desconocida Identificar interacciones entre proteínas Predicción estructura secundaria Determinar presencia de motivos Predicción estructura terciaria Determinar función de la proteína Objetivos: Proteómica: Objetivos

© Copyright Ebiointel,SL 2006 Comparar proteomas distintos

© Copyright Ebiointel,SL 2006 Comparar proteomas distintos

© Copyright Ebiointel,SL 2006 Tecnicas experimentales: -Secuenciación de novo de péptidos -Microchips de proteínas -MALDI-TOF -geles bidimensionales combinadas con la bioinformatica Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificación de una proteína a partir de sus características fisicoquímicas y a partir de una digestión enzimática AnalysisWhole Protein Molecular Weight m.w. Length501 Isoelectric Point5.40 Charge at pH AnalysisWhole Protein Molecular Weight m.w. Length501 Isoelectric Point5.40 Charge at pH Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 N. count % by weight % by frequency Acidic (DE) Basic (KR) Polar (NCQSTY) Hydrophobic (AILFWV) N. count % by weight % by frequency A Ala C Cys D Asp E Glu F Phe G Gly H His I Ile K Lys L Leu M Met N Asn P Pro Q Gln N. count % by weight % by frequency R Arg S Ser T Thr V Val W Trp Y Tyr B Asx Z Glx X Xxx Ter Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Hydroxylamine - 4 cuts Position LengthWeightpI1/2 life HPLC rtFragment >20h81.24MAPALHWLL...GSFVEMVDN m85.54LRGKSGQGY...TESDKFFIN >20h88.02GSNWEGILG...VIIVRVEIN >20h57.76GQDLKMDCK...SIVDSGTTN m96.18LRLPKKVFE...FADDISLLK Digestión enzimática Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Digestión enzimática Identificar una proteína desconocida

© Copyright Ebiointel,SL Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 MRVLLVALALLALAASATSTHTSGGCGCQPPPPVHLPPPVHLPPPVHLPP PVHLPPPVHLPPPVHLPPPVHVPPPVHLPPPPCHYPTQPPRPQPHPQPHPC PCQQPHPSPCQLQGTCGVGSTPILGQCVEFLRHQCSPTATPYCSPQCQSL RQQCCQQLRQVEPQHRYQAIFGLVLQSILQQQPQSGQVAGLLAAQIAQQ LTAMCGLQQPTPCPYAAAGGVPH Proteína problema (obtenida directamente del banco de datos) Purificada de maiz (Zea mays) Pm: ¿? pI: ¿? Composicio de aa: ¿? Masa molecular de los peptidos por digestión con tripsina: ¿? Purificada de maiz (Zea mays) Pm: ¿? pI: ¿? Composicio de aa: ¿? Masa molecular de los peptidos por digestión con tripsina: ¿?

© Copyright Ebiointel,SL 2006

Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 PROTEINA PROBLEMA: Purificada de maiz (Zea mays) Pm: pI: 8.40 Composicio de aa: Ala (A) % Arg (R) 6 2.7% Asn (N) 0 0.0% Asp (D) 0 0.0% Cys (C) % Gln (Q) % Glu (E) 2 0.9% Gly (G) % His (H) % Ile (I) 4 1.8% Leu (L) % Lys (K) 0 0.0% Met (M) 2 0.9% Phe (F) 2 0.9% Pro (P) %Ser (S) % Thr (T) % Trp (W) 0 0.0% Tyr (Y) 4 1.8% Val (V) % Masa molecular de los peptidos por digestión con tripsina: PROTEINA PROBLEMA: Purificada de maiz (Zea mays) Pm: pI: 8.40 Composicio de aa: Ala (A) % Arg (R) 6 2.7% Asn (N) 0 0.0% Asp (D) 0 0.0% Cys (C) % Gln (Q) % Glu (E) 2 0.9% Gly (G) % His (H) % Ile (I) 4 1.8% Leu (L) % Lys (K) 0 0.0% Met (M) 2 0.9% Phe (F) 2 0.9% Pro (P) %Ser (S) % Thr (T) % Trp (W) 0 0.0% Tyr (Y) 4 1.8% Val (V) % Masa molecular de los peptidos por digestión con tripsina: Identificar una proteína desconocida

© Copyright Ebiointel,SL 2006 Identificar interacciones entre proteínas Interacciones descritas en el proteoma de levadura

© Copyright Ebiointel,SL Identificar interacciones entre proteínas

© Copyright Ebiointel,SL 2006 Identificar interacciones entre proteínas