Post on 02-Feb-2016
Movimiento de Atraccióny
Formación del Complejo
Long-range interactions (global charges)
-+
+++ ---
Short-range interactions (local charges)
Reorientation
Collision
Sliding along themembrane surface
Free diffusion
Interaction between Plastocyanin and PSI
Sliding along themembrane surface
Collision
Reorientation
Free diffusion
Electrostatic Protein-Protein Interactions
Long-range (global charges)Monopole-monopole interactions
Short-range (local charges)Dipole-dipole interactionsMonopole-dipole interactions
Dipole-Dipole Interaction of Plastocyanin with PSI
--
+ -
--+ -
--+
-
--
+ -
--
+
-
Heme P62H87
Y83
Cytochrome c6 Plastocyanin
Frazão et al. (1995) Structure 3, 1159-1169
Cytochrome c6 Plastocyanin
R67
Frazão et al. (1995) Structure 3, 1159-1169
Cytochrome c6 Plastocyanin
P62Heme H87
Frazão et al. (1995) Structure 3, 1159-1169
Cytochrome f
Plastocyanin
Frazão et al. (1995) Structure 3, 1159-1169
Mutagénesis Dirigida
Análisis Funcional de las Interacciones entre Proteínas
Lys33(35)His87(92)
Leu12(14)
Arg88(93)
Tyr83(88)
Lys58(62)
His57(61)
Asp49(54)AnabaenaPlastocyanin
Molina-Heredia et al. (2001) J. Biol. Chem. 276, 601-605
WTK33E K58A D49K
+-
Reactivity towards photosystem I
Surface electrostatic potential
Anabaena Plastocyanin
Con
stan
te d
e ve
loci
dad
Fuerza iónica
Efecto de la fuerza iónica sobre las interacciones entre proteínas
Val25
Arg64Lys66
Asp72
Lys62Lys29
hydrophobicpatch
Cytochrome c6 from AnabaenaTarget residues for site-directed mutagenesis
chargedarea
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
Cytochrome c6Plastocyanin
Anabaena
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 Redoxy
Disociación del Complejo
Plastocyanin-PSI Complex
E. Myshkin et al. (2002) Biophys J. 82, 3305
Cyt c6Pc
PS I
b6f
PSI-driven Electron Transfer
Fd
light
Green algaC. reinhardtii
CyanobacteriumPhormidium laminosum
Cytochrome f
Higher plantBrassica rapa
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
Ubbink et al. (1998) Structure 6, 323-335
Electron transfer pathway from cytochrome f to plastocyanin
Crowley et al. (2002) submitted
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
Complejo INADH-quinonaóxido-reductasa Complejo II
Succinato-quinonaóxido-reductasa
Complejo IIIQuinol-citocromo cóxido-reductasa
Complejo IVCitocromo c-O2
óxido-reductasa
citocromo c
Cadena respiratoria mitocondrial
Complejo III (o citocromo bc1)Quinol-citocromo c óxido-reductasa
Cyt c
Cyt c1
Cyt c
Cyt c1
Interacción entre el complejo bc1 (inmerso en la membrana) y el citocromo c (soluble)
Interacción entre los citocromos f y c
Cyt c1
Cyt c
Cyt c1
Cyt c
Hemo c
Cyt c1
Hemo c1Cyt c
Zona de contacto entre los hemos f y c
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
Centro de Investigaciones
Científicas
Isla de la Cartuja
Sevilla
Universidad de Sevilla
http://transientprot.ibvf.csic.es
Workshop on Weak Protein-Protein Interactions
Seville, SpainJuly 9-13, 2003
Target residues for mutagenesisTarget residues for mutagenesisof cytochrome of cytochrome cc66
AnabaenaSynechocystisMonoraphidium
AnabaenaSynechocystisMonoraphidium
AnabaenaSynechocystisMonoraphidium
1
1
1
31
31
32
59
59
62
303031
585861
868589
D L A LG K A V F D GN C A AC H AGGGN N V I P D H T
L K K AD L E K Y - - GMY S A E A I I AQ V T N G KN - AML KMAD L E AN - - G KN S V A A I V AQ I T N G - N G AML Q K A A I E Q F L D G KN S V A A I V AQ I T N G - N G AM
P A F G L KK P E Q I E V A A Y V L G K AD A
- AD S V N G A K I F S AN C AS C H AGG KN L Q AQ T- AD L AH G K A I F AGN C A AC H N GG L N A I N P S K TE A
DWKP G KG I S DDS DD ME D V A A Y V L D Q A E KGWP G F KG R I S D S D ME D V A A Y V L D Q A - AGN KW
FFRRRR
KKVV
KK DD
Molina-Heredia et al. (1999) J. Biol. Chem. 274, 33565 De la Cerda et al. (1999) J. Biol. Chem. 274, 13292
0 100 200Cytochrome c6 (M)
kob
s x 1
0-4 (
s-1)
0
2
4
6
Anabaena
Molina-Heredia et al. (1999) JBC 274, 33565
WT
D72K
V25AV25EK29HK62ER64EK66E
WT
0 100 200 300
0
2
4
8
6
Cytochrome c6 (mM)
kob
s x 1
0-4 (
s-1)
Anabaena
Molina-Heredia et al. (1999) JBC 274, 33565
V25A
D72K
Synechocystis Plastocyanin
De la Cerda et al. (1997) Biochemistry 36, 10125
0.2 0.3 0.4 0.5 0.6
Square root of ionic strength (M1/2)
0.5
1.0
1.5
2.0
0k o
bs x
10-
3 (s
-1)
WT
D47R
D47A
Synechocystis Plastocyanin
De la Cerda et al. (1997) Biochemistry 36, 10125
0.2 0.3 0.4 0.5 0.6
Square root of ionic strength (M1/2)
0.5
1.0
1.5
2.0
0k o
bs x
10-
3 (s
-1)
D44A
D44K
WT
Synechocystis Plastocyanin
De la Cerda et al. (1997) Biochemistry 36, 10125
0.2 0.3 0.4 0.5 0.6
Square root of ionic strength (M1/2)
0.5
1.0
1.5
2.0
0k o
bs x
10-
3 (s
-1)
WT
D44R/D47R
D44A/D47A
ReducedOxidised
Cytochrome c6 from Monoraphidium
ReducedOxidised
Cytochrome c6 from Monoraphidium
Bioreactor forgrowing cells