BIOQUÍMICA

BÁSICA
1000042-1
I-2019

FOSFORILACIÓN OXIDATIVA
 LA IMPORTANCIA DE LA
FOSFORILACIÓN OXIDATIVA

EVENTO BIOQUÍMICO DONDE

CULMINA LA OBTENCIÓN DE
ENERGÍA QUÍMICA EN LOS
ORGANISMOS AEROBIOS
 LOS 3 EVENTOS FUNDAMENTALES DE LA
FOSFORILACIÓN OXIDATIVA

1- TRANSPORTAR ELECTRONES A LO
LARGO DE UNA CADENA
TRANSPORTADORA

2- ESTABLER UN GRADIENTE DE
PROTONES A TRAVÉS DE UNA
MEMBRANA BIOLÓGICA

3- ACOPLAR EL FLUJO DE PROTONES

A FAVOR DE SU GRADIENTE CON LA
SÍNTESIS DE ATP
 ¿DÓNDE OCURRE LA FOSFORILACIÓN OXIDATIVA?

1- EN LA
MITOCONDRIA
DE ORGANISMOS
EUCARIOTAS

2- EN LA MEMBRANA
CELULAR
DE ORGANISMOS
PROCARIOTAS
MOLÉCULAS TRANSPORTADORAS DE ELECTRONES

IONES

MODALIDADES DE TRANSFERENCIA DE
1-NAD
HIDRURO
2-FAD
ÁTOMOS DE
HIDRÓGENO

ELECTRONES
3-UBIQUINONA

4-CITOCROMOS
(CONTIENEN GRUPOS ELECTRONES
PROSTÉTICOS TIPO HEMO) DESNUDOS
Cambia estado de
5-PROTEÍNAS CON oxidación:
CENTROS DE Fe3+ a Fe2+
HIERRO-AZUFRE
 MOLÉCULAS TRANSPORTADORAS DE ELECTRONES
NAD 13.3 Biological Oxidation-Reduction Reactions 513

O O O
H B H H B H H B
C 2 e$
? C ? CH
H H
NH 2 NH 2 or NH 2 ! H!
!
N 2 H! N N
O CH 2 O A A
R A side R B side
H H
OP P O O H$ N AD H
H
(r edu ced)

O OH OH
NH 2
N
OP P O O $ N Aden in e
1.0
Oxidized
N N (NAD!)
O CH 2 O 0.8

Absor ba n ce
H H 0.6
H H
!
N AD Redu ced
(oxidized) 0.4
OH OH (NADH )

In NADP ! t h is h ydr oxyl gr ou p 0.2

is est er ified wit h ph osph a t e.
0.0
(a ) 220 240 260 280 300 320 340 360 380
Wa velen gt h (n m )
(b)

FIGURE 13–15 NAD and NADP. (a) Nicotinamide adenine dinu- tra of NAD! and NADH. Reduction of the nicotinamide ring produces
cleotide, NAD!, and its phosphorylated analog NADP! undergo re- a new, broad absorption band with a maximum at 340 nm. The pro-
 MOLÉCULAS TRANSPORTADORAS DE ELECTRONES
516 Chapter 13 Principles of Bioenergetics
FAD
isoa lloxa zin e r in g
O H O" H O
!
CH 3 N H !e! " CH 3 N !
H !e "
CH 3 N
NH • NH NH

CH 3 N N O CH 3 N N O CH 3 N N O
CH 2 R R H
H COH FADH • (F MNH • ) FADH 2 (F MNH 2 )
F MN (sem iqu in on e) (fu lly r edu ced)
H COH
H COH
CH 2
FAD
O
"
O P O
O
NH 2
"
O P O N
N
O
N N
CH 2 O
FIGURE 13–18 Structures of oxidized and reduced FAD and FMN.
H H FMN consists of the structure above the dashed line on the FAD (ox-
H H
idized form). The flavin nucleotides accept two hydrogen atoms (two
OH OH electrons and two protons), both of which appear in the flavin ring
F la vin a den in e din u cleot ide (FAD) a n d system. When FAD or FMN accepts only one hydrogen atom, the semi-
fla vin m on on u cleot ide (F MN) quinone, a stable free radical, forms.
MOLÉCULAS TRANSPORTADORAS DE ELECTRONES
UBIQUINONA
MOLÉCULAS TRANSPORTADORAS DE ELECTRONES
CITOCROMOS CON GRUPOS PROSTÉTICOS HEMO
MOLÉCULAS TRANSPORTADORAS DE ELECTRONES
PROTEÍNAS CON CENTROS DE HIERRO-AZUFRE
 inorganic S atoms are counted. For example, in the 2Fe-2S center
(b), each Fe ion is actually surrounded by four S atoms.) The exact

¿EN QUÉ ORDEN SE TRANSFIEREN LOS ELECTRONES?

standard reduction potential of the iron in these centers depends on
the type of center and its interaction with the associated protein.

DESDE MOLÉCULAS CON POTENCIALES DE REDUCCIÓN BAJOS

First, the standard reduction potentials of the in- Q → cytochrome b → cytochrome c1 → cytochrome
dividual electron carriers have been determined ex- c → cytochrome a → cytochrome a3 → O2. Note, how-
HACIA MOLÉCULAS CON POTENCIALES ALTOS
perimentally (Table 19–2). We would expect the carri- ever, that the order of standard reduction potentials is
ers to function in order of increasing reduction not necessarily the same as the order of actual reduc-
potential, because electrons tend to flow spontaneously tion potentials under cellular conditions, which depend
POTENCIAL DE REDUCCIÓN:
from carriers of lower E!" to carriers of higher E!". The on the concentration of reduced and oxidized forms
MEDIDA DE LA AFINIDAD DE UN PAR REDOX POR LOS ELECTRONES
order of carriers deduced by this method is NADH → (p. 510). A second method for determining the sequence

TABLE 19–2 Standard Reduction Potentials of Respiratory Chain and Related Electron Carriers
Redox reaction (half-reaction) E !" (V)
2H# # 2e$ 8n H2 $ 0.414
NAD# # H# # 2e$ 8n NADH $ 0.320
NADP# # H# # 2e$ 8n NADPH $ 0.324
NADH dehydrogenase (FMN) # 2H# # 2e$ 8n NADH dehydrogenase (FMNH2) $ 0.30
Ubiquinone # 2H# # 2e$ 8n ubiquinol 0.045
Cytochrome b (Fe3# ) # e$ 8n cytochrome b (Fe2# ) 0.077
Cytochrome c1 (Fe3# ) # e$ 8n cytochrome c1 (Fe2# ) 0.22
Cytochrome c (Fe3# ) # e$ 8n cytochrome c (Fe2# ) 0.254
Cytochrome a (Fe3# ) # e$ 8n cytochrome a (Fe2# ) 0.29
Cytochrome a3 (Fe3# ) # e$ 8n cytochrome a3 (Fe2# ) 0.35
%%O #
1
2 2 2H# # 2e$ 8n H2O 0.8166
¿EN QUÉ ORDEN SE TRANSFIEREN LOS ELECTRONES?

DESDE MOLÉCULAS CON POTENCIALES DE REDUCCIÓN BAJOS

HACIA MOLÉCULAS CON POTENCIALES ALTOS
Energy Metabolism 143

A. Redox systems of the respiratory chain

∆G' E'
FMN/FMNH2
(kJ · mol–1 ) (V)
F F
NAD
–0.4
N A
I CoQ Hemes b
(ubi-
0 quinone) 3+ 2+
–0.2 N A O

NADH
+H III
0 O Cyto-
chrome C
OH Heme a
Cu2
+0.2 Cu 3+ 2+
Fe/S centers 3+

OH
IV
2+
+0.4
Fe/S center Cu2
Cu
3+ 2+
+0.6
Heme c1
3+ 2+
∆G = –n · F · ∆E O
–220 +0.8
Heme a3 O2

B. ATP synthase
vestigators have determined the entire sequence; it is composed of 42 different polypeptide chains, including
the same as deduced in the first two approaches. an FMN-containing flavoprotein and at least six iron-

LA CADENA TRANSPORTADORA DE ELECTRONES

Electron Carriers Function in Multienzyme Complexes
sulfur centers. High-resolution electron microscopy
shows Complex I to be L-shaped, with one arm of the L

PRESENTA 4 COMPONENTES PROTEICOS

The electron carriers of the respiratory chain are or-
in the membrane and the other extending into the ma-
trix. As shown in Figure 19–9, Complex I catalyzes two
ganized into membrane-embedded supramolecular simultaneous and obligately coupled processes: (1) the

TABLE 19–3 The Protein Components of the Mitochondrial Electron-Transfer Chain

Enzyme complex/protein Mass (kDa) Number of subunits* Prosthetic group(s)
I NADH dehydrogenase 850 43 (14) FMN, Fe-S
II Succinate dehydrogenase 140 4 FAD, Fe-S
III Ubiquinone cytochrome c oxidoreductase 250 11 Hemes, Fe-S
Cytochrome c† 13 1 Heme
IV Cytochrome oxidase 160 13 (3–4) Hemes; CuA, CuB

*
Numbers of subunits in the bacterial equivalents in parentheses.
†
Cytochrome c is not part of an enzyme complex; it moves between Complexes III and IV as a freely soluble protein.

DADAS SUS CARÁCTERÍSTICAS BIOQUÍMICAS, LOS 4 COMPONENTES

SE PUEDEN AISLAR Y ESTUDIAR POR SEPARADO EN EL LABORATORIO
 SEPARACIÓN DE LOS
COMPONENTES PROTEICOS DE
LA CADENA TRANSPORTADORA
DE ELECTRONES

LA ATP SINTASA SE DENOMINA

COMÚNMENTE COMO COMPLEJO V

EN EL LABORATORIO, SÓLO PROCEDE
CON LA HIDRÓLISIS DE ATP
EL COMPLEJO I: NADH DESHIDROGENASA
EL COMPLEJO II: SUCCINATO DESHIDROGENASA

Función del grupo hemo b:

Evitar “goteo” de electrones

El complejo II también
participa en el ciclo de Krebs
EL COMPLEJO III: CITOCROMO bc1
EL COMPLEJO III: CITOCROMO bc1
EL COMPLEJO IV: CITOCROMO OXIDASA
EL COMPLEJO IV: CITOCROMO OXIDASA
EL FLUJO DE ELECTRONES A LO LARGO DE LA
CADENA TRANSPORTADORA

QH2 y Cyt c difunden de un sitio a otro

A A 3.6.1.34
P P P
nH P P P

Mr >400 kDa, >20 subunits

EL FLUJO DE ELECTRONES A LO LARGO DE LA

1/2 O
O
2H
Electron flow 2
2

CADENA TRANSPORTADORA
H O +0.8 Proton flow
2

B. Organization

Outer
mitochon-
drial
membrane
Inter-
membrane
4H 2H space
4H
II
Inner
I mitochon-
Cyto- ?
Q chrome C ∆p drial
III IV V membrane

2e 4H 2-4 H
4H 1/2 O2 Matrix
O2 3 P space
2H
N A N A A A
3 P P
3 P P P

ATP
Tricarboxylic H2O 10 H
NAD
NADH+H acid cycle
β-Oxidation
EL ESTABLECIMIENTO DE UN GRADIENTE
ELECTRO-QUÍMICO
I is not yet known, but probably involves a Q cycle similar to that in succinate. The path of electron transfer from the
Complex III in which QH2 participates twice per electron pair (see succinate-binding site to FAD, then through the Fe-S

INHIBIDORES DE LA CADENA TRANSPORTADORA DE

Fig. 19–12). Proton flux produces an electrochemical potential across
the inner mitochondrial membrane (N side negative, P side positive),
centers to the Q-binding site, is more than 40 Å long,
but none of the individual electron-transfer distances

ELECTRONES
which conserves some of the energy released by the electron-transfer
reactions. This electrochemical potential drives ATP synthesis.
exceeds about 11 Å—a reasonable distance for rapid
electron transfer (Fig. 19–10).

TABLE 19–4 Agents That Interfere with Oxidative Phosphorylation or Photophosphorylation

Type of interference Compound* Target/mode of action
Inhibition of electron transfer Cyanide ⎫
Inhibit cytochrome oxidase
⎬
Carbon monoxide ⎭
Antimycin A Blocks electron transfer from cytochrome b to cytochrome c1
Myxothiazol ⎫
Rotenone ⎪
⎬ Prevent electron transfer from Fe-S center to ubiquinone
Amytal ⎪
Piericidin A ⎭
DCMU Competes with QB for binding site in PSII
Inhibition of ATP synthase Aurovertin Inhibits F1
Oligomycin ⎫
⎬ Inhibit Fo and CFo
Venturicidin ⎭
DCCD Blocks proton flow through Fo and CFo
Uncoupling of phosphorylation FCCP ⎫
⎬ Hydrophobic proton carriers
from electron transfer DNP ⎭
Valinomycin K! ionophore
Thermogenin In brown fat, forms proton-conducting pores in inner mitochondrial
membrane
Inhibition of ATP-ADP exchange Atractyloside Inhibits adenine nucleotide translocase

*
DCMU is 3-(3,4-dichlorophenyl)-1,1-dimethylurea; DCCD, dicyclohexylcarbodiimide; FCCP, cyanide-p-trifluoromethoxyphenylhydrazone; DNP,
2,4-dinitrophenol.
EN OCASIONES A LA UBIQUINONA SE LE
ESCAPAN ELECTRONES
 LA SÍNTESIS DE ATP:
EL MODELO ELECTRO-QUÍMICO

LOS PROTONES FLUYEN PASIVAMENTE DE REGRESO A LA MATRIZ MITOCONDRIAL

APROVECHANDO EL GRADIENTE ELECTRO-QUÍMICO
LA ATP-SINTASA (COMPLEJO V)
LA ATP-SINTASA (COMPLEJO V)
 Heme c1
3+ 2+
∆G = –n · F · ∆E O
–220 +0.8
CATÁLISIS ROTACIONAL
Heme a 3
O2

B. ATP synthase

Inside: A
P P P
matrix 2 Formation of ATP

A
P P P
H2O
Outside: H2O
A
P P P
Inter-
membrane H H
space outside outside

α β α
δ
β α β
F1

A
P
H
P P
inside
b2 H
ε A A
P P P
P P P

γ
A
P P P

a F0 A
A H2O P P P H2O
P P P A A
P P P
H2O
P P P H2O

H2O

1 Binding of ADP and Pi 3 Release of ATP

H H
outside

1. Structure and location 2. Catalytic cycle

CATÁLISIS ROTACIONAL
LA MITOCONDRIA COMO UNA CENTRAL
ENERGÉTICA
 PREMIOS NOBEL

MODELO ESTRUCTURA CATALISIS

QUIMIO-OSMÓTICO ATP-SINTASA ROTACIONAL
TRANSPORTE DE MOLÉCULAS A EXPENSAS DEL
GRADIENTE DE PROTONES
¿CÓMO TRANSPORTAR EL NADH DEL CITOSOL
HACIA LA MATRIZ MITOCONDRIAL?
1-MEDIANTE LA LANZADERA MALATO-ASPARTATO
¿CÓMO TRANSPORTAR EL NADH DEL CITOSOL
HACIA LA MATRIZ MITOCONDRIAL?
2-MEDIANTE LA LANZADERA DE GLICEROL 3 FOSFATO
 rison, glycolysis under anaerobic conditions mass-action ratio returns to its normal hig
ermentation) yields only 2 ATP per glucose. which point respiration slows again. The rat
BALANCE ENERGÉTICO
e evolution of oxidative phosphorylation pro- tion of cellular fuels is regulated with such
emendous increase in the energy efficiency of and precision that the [ATP]/([ADP][Pi]) ratio
m. Complete oxidation to CO2 of the coenzyme only slightly in most tissues, even during ex
ve of palmitate (16:0), which also1 NADH= 2,5 ATPs
occurs in ations in energy demand. In short, ATP is f
hondrial matrix, yields 108 ATP per palmitoyl- as fast as it is used in energy-requiring cellula
1 FADH2= 1,5 ATPs

TABLE 19–5 ATP Yield from Complete Oxidation of Glucose

Process Direct product Final ATP
Glycolysis 2 NADH (cytosolic) 3 or 5 *
2 ATP 2
Pyruvate oxidation (two per glucose) 2 NADH (mitochondrial matrix) 5
Acetyl-CoA oxidation in citric acid cycle 6 NADH (mitochondrial matrix) 15
(two per glucose) 2 FADH2 3
2 ATP or 2 GTP 2
Total yield per glucose 30 or 32

*
The number depends on which shuttle system transfers reducing equivalents into the mitochondrion.
 pyruvate kinase ADP

ATP, NADH

REGULACIÓN DE LA FOSFORILACIÓN OXIDATIVA pyruvate

Pyruvate
AMP, ADP, NAD!
dehydrogenase
complex ATP, NADH
718 Chapter 19 Oxidative Phosphorylation and Photophosphorylation Acetyl-CoA
ADP
citrate synthase
ATP, NADH
Glucose FIGURE 19–31 Regulation of the ATP-producing pathways. This di-
Citrate
agram shows the interlocking regulation of glycolysis, pyruvate oxi-
hexokinase Pi dation, the citric acid cycle, and oxidative phosphorylation by the rel-
ative concentrations of ATP, ADP, and AMP, and by NADH. High [ATP]
ADP
isocitrate
Glucose 6-phosphate (or low [ADP] and [AMP]) produces low rates of glycolysis, pyruvate
dehydrogenase ATP
oxidation, acetate oxidation via the citric acid cycle, and oxidative
phosphorylation. All four pathways"-Ketoglutarate
are accelerated when the use of
AMP, ADP Citric
ATP andacid -ketoglutarate
the formation of ADP, AMP, and Pi increase. The interlock-
"
phosphofructokinase-1
ATP, citrate cycle dehydrogenase ATP, NADH
ing of glycolysis and the citric acid cycle by citrate, which inhibits
glycolysis, supplements the action of the adenine nucleotide system.
Succinyl-CoA
Fructose 1,6-bisphosphate
Glycolysis In addition, increased levels of NADH and acetyl-CoA also inhibit the
oxidation of pyruvate to acetyl-CoA, and a high [NADH]/[NAD!] ra-
multistep tio inhibits the dehydrogenase reactions of the citric acid cycle (see
multistep
Fig. 16–18).

Phosphoenolpyruvate
ADP
pyruvate kinase Oxaloacetate
ATP, NADH

Pyruvate
ATP-Producing Pathways Are Coordinately
ADP, Pi Regulated
1
pyruvate
AMP, ADP, NAD! The NADHpathways have interlocking
major catabolic
Oxidative 2 O2 and
dehydrogenase
phosphory-
concerted Respiratory chain
regulatory mechanisms that allow them to
complex ATP, NADH lation
function togetherNAD H2O
!
in an economical and self-regulating
Acetyl-CoA
manner to produce ATPADP and! Pbiosynthetic
i ATP precursors.
ADP
citrate synthase The relative concentrations of ATP and ADP control not
ATP, NADH only the rates of electron transfer and oxidative phos-
LA TERMOGENINA Y LA GENERACIÓN DE CALOR
BIBLIOGRAFÍA
A. Reactive oxygen species B. Biological antioxidants

Molecular Quinols α-Tocopherol (vitam

O2 and enols Ubiquinol (coenzym
oxygen
Ascorbic acid (vitam
e
a Carotenoids β-Carotin
Dispro- Lycopin
portionation
Others Glutathione
Bilirubin
Superoxide 1
radical O2
1. Examples

e Glu Cys
b
H O H O
Hydrogen
peroxide OOC C (CH2)2 C N C C N CH

2 NH3 H CH2 H
O2 H 2 O2 2 2 Glu
SH
2 (GS
Hydroxy e 2H Oxidation Reduction
radical c
H O H O
2
OH O O H2 O OOC C (CH2)2 C N C C N CH
NH3 H CH2 H
d Water
H 2H S Glutat
disulfid
S
e NH3 H CH2 H
OOC C (CH2)2 C N C C N CH
H O H O
1 Superoxide dismutase 2 Catalase
1.15.1.1 1.11.1.6 2. Glutathione

C. Erythrocyte metabolism