Movimiento de Atracción Formación del Complejo y Formación del Complejo

- + - Long-range interactions (global charges) Short-range interactions (local charges)

Collision Reorientation Sliding along the membrane surface Free diffusion

Electrostatic Protein-Protein Interactions Long-range (global charges) Monopole-monopole interactions Short-range (local charges) Dipole-dipole interactions Monopole-dipole interactions

D i p o l e - I n t r a c f P s y w h S - + - + - + - + - +

Cytochrome c6 Plastocyanin H87 Heme P62 Y83 Frazão et al. (1995) Structure 3, 1159-1169

Cytochrome c6 Plastocyanin R67 Frazão et al. (1995) Structure 3, 1159-1169

Cytochrome c6 Plastocyanin Heme H87 P62 Frazão et al. (1995) Structure 3, 1159-1169

Plastocyanin Cytochrome f Frazão et al. (1995) Structure 3, 1159-1169

Análisis Funcional de las Interacciones entre Proteínas Mutagénesis Dirigida Análisis Funcional de las Interacciones entre Proteínas

Anabaena Plastocyanin Lys33(35) His87(92) Lys58(62) Leu12(14) Arg88(93) Tyr83(88) Lys58(62) His57(61) Asp49(54) Anabaena Plastocyanin Molina-Heredia et al. (2001) J. Biol. Chem. 276, 601-605

- + Anabaena Plastocyanin K33E K58A WT D49K Surface electrostatic potential Reactivity towards photosystem I

Efecto de la fuerza iónica sobre las interacciones entre proteínas Constante de velocidad Fuerza iónica

Cytochrome c6 from Anabaena Target residues for site-directed mutagenesis Lys29 Lys62 hydrophobic patch charged area Val25 Asp72 Arg64 Lys66 Molina-Heredia et al. (1999) J. Biol. Chem. 274, 33565-70

Anabaena Cytochrome c6 WT V25E K62E R64E K66E D72K Molina-Heredia et al. (1999) J. Biol. Chem. 274, 33565-70

Anabaena Plastocyanin Molina-Heredia et al. (2001) J. Biol. Chem. 276, 601-605

Anabaena Molina-Heredia et al. (2001) J. Biol. Chem. 276, 601-605

Anabaena Plastocyanin Cytochrome c6 Molina-Heredia et al. (2001) J. Biol. Chem. 276, 601-605

De la Rosa et al. (2002) Bioelectrochemistry 55, 41-45

Orientación de los Centros Redox Disociación del Complejo y Disociación del Complejo

Plastocyanin-PSI Complex E. Myshkin et al. (2002) Biophys J. 82, 3305

PSI-driven Electron Transfer Fd light PS I b6f Cyt c6 Pc

Cytochrome f Green alga C. reinhardtii Cyanobacterium Phormidium laminosum Higher plant Brassica rapa Cytochrome f

Electrostatic complex between cytochrome f and plastocyanin Ubbink et al. (1998) Structure 6, 323-335

Electrostatic complex between cytochrome f and plastocyanin Ubbink et al. (1998) Structure 6, 323-335

Electron transfer pathway from cytochrome f to plastocyanin Ubbink et al. (1998) Structure 6, 323-335

Chemical shift map of Ana-cytc6 in the presence of cytf Residues are coloured according to increasing Bind from blue to red Prolines and the haem group are in grey and green, respectively Crowley et al. (2002) submitted

Cadena respiratoria mitocondrial Complejo I NADH-quinona óxido-reductasa Complejo II Succinato-quinona Complejo III Quinol-citocromo c Complejo IV Citocromo c-O2 citocromo c

Complejo III (o citocromo bc1) Quinol-citocromo c óxido-reductasa

Interacción entre el complejo bc1 (inmerso en la membrana) y el citocromo c (soluble) Cyt c Cyt c1 Cyt c Cyt c1

Interacción entre los citocromos f y c Cyt c1 Cyt c

Zona de contacto entre los hemos f y c Cyt c1 Hemo c Cyt c Hemo c1

Main conformational changes between the oxidized (red) and reduced (blue) forms of Monoraphidium cytochrome c6 De la Rosa et al. (2002) Bioelectrochemistry 55, 41-45

Protein-Protein Interactions Centro de Investigaciones Científicas Isla de la Cartuja Sevilla Universidad de Sevilla Workshop on Weak Protein-Protein Interactions Seville, Spain July 9-13, 2003 http://transientprot.ibvf.csic.es

Target residues for mutagenesis of cytochrome c6 Anabaena 1 31 32 59 62 30 58 61 86 85 89 D L A G K V F N C H I P T E Y - M S Q W R Synechocystis Monoraphidium Anabaena Synechocystis Monoraphidium Anabaena Synechocystis Monoraphidium Molina-Heredia et al. (1999) J. Biol. Chem. 274, 33565 De la Cerda et al. (1999) J. Biol. Chem. 274, 13292

Anabaena 100 200 Cytochrome c6 (mM) kobs x 10-4 (s-1) 2 4 6 D72K WT 100 200 Cytochrome c6 (mM) kobs x 10-4 (s-1) 2 4 6 Anabaena Molina-Heredia et al. (1999) JBC 274, 33565 WT D72K V25A V25E K29H K62E R64E K66E

Anabaena 100 200 300 2 4 8 6 Cytochrome c6 (mM) kobs x 10-4 (s-1) D72K WT 100 200 300 2 4 8 6 Cytochrome c6 (mM) kobs x 10-4 (s-1) Anabaena Molina-Heredia et al. (1999) JBC 274, 33565 D72K V25A

Synechocystis Plastocyanin De la Cerda et al . (1997) Biochemistry 36, 10125

Synechocystis Plastocyanin De la Cerda et al . (1997) Biochemistry 36, 10125

Synechocystis Plastocyanin De la Cerda et al . (1997) Biochemistry 36, 10125

Cytochrome c6 from Monoraphidium Reduced Oxidised

Cytochrome c6 from Monoraphidium Reduced Oxidised

Bioreactor for growing cells