Week 12 - Biology 112

Most of the energy captured from glucose oxidation is in

the form of reduced electron carriers (NADH and FADH
2
)
Three mechanisms of ATP synthesis
Light-independent synthesis - catabolism
1. Substrate-level phosphorylation
2. Oxidative phosphorylation
Light-dependent synthesis - photosynthesis
3. Photophosphorylation
Oxidative phosphorylation
Depends on the function of the electron transport chain
(ETC) and the ATP synthase enzyme
Eukaryotic mitochondrion Bacterial cell
Outer
membrane
Intermembrane
space
Matrix
Inner
membrane
Cell wall
Cytoplasmic
membrane
Periplasm Cytoplasm
Eukaryotes
Inner mitochondrial membrane
Bacteria
Cytoplasmic membrane
Location
Mitochondria: the sites of oxidative phosphorylation in
eukaryotic cells.
Remember, plant cells have mitochondria, too!
!"#$ &'() *'&&
+,-"*."$/#,(
!.&"#"0&(1-
Modes of catabolism
Respiration
Aerobic
Anaerobic
Fermentation
Cell making ALL of its
ATP by substrate level
phosphorylation
Some bacteria use other
TEAs
eg. NO
3
-
, SO
4
-2

Cell making MOST of
its ATP by oxidative
phosphorylation
TEA = O
2
TEA = O
2
Electron transport chain
2$$'#
3,-"*."$/#,(&
3'34#($'
"# *5-"0&(13,*
3'34#($'
+(-#,6 "#
*5-"0&(13
2$-'# 3'34#($'
10(*'
"# 0'#,0&(13
Mostly large membrane protein complexes
One lipid component Q
Protein complexes have bound electron
carrying groups that undergo oxidation-
reduction.
2$$'#
3,-"*."$/#,(&
3'34#($'
"# *5-"0&(13,*
3'34#($'
+(-#,6 "#
*5-"0&(13
2$-'# 3'34#($'
10(*'
"# 0'#,0&(13
Example: Cytochrome c
has a bound Fe-atom
Goes through a redox cycle = 2 redox reactions
7'8(-"3
7#"3
!"30&'6 222
9"
!"30&'6 2:
2$$'#
3,-"*."$/#,(&
3'34#($'
"# *5-"0&(13,*
3'34#($'
+(-#,6 "#
*5-"0&(13
2$-'# 3'34#($'
10(*'
"# 0'#,0&(13
Oxidative phosphorylation is an oxidation-reduction (redox)-
based process
Electron donors (carriers) are oxidized: NADH, FADH
2

(Terminal) electron acceptor O
2
is reduced
2$$'#
3,-"*."$/#,(&
3'34#($'
"# *5-"0&(13,*
3'34#($'
+(-#,6 "#
*5-"0&(13
2$-'# 3'34#($'
10(*'
"# 0'#,0&(13
As electrons from the carriers pass through the chain, some
of the protein complexes capture the energy from their redox
cycles and use it to actively transport H
+
across the
membrane
The transported H+ comes from matrix or cytoplasmic water.
Remember water = H
2
O + HO
-
+ H
+

Transport of H
+
creates a H
+
concentration difference (=
gradient) across the membrane.
Transport of H
+
without transport of HO
-
(= separating
charge)

creates an electrical difference (= potential, gradient)
across the membrane
!"$*'$-#(-,"$
;#(/,'$-
<
=&'*-#,*(&
;#(/,'$-
>
=&'*-#"*.'3,*(&
;#(/,'$-
Final step: Synthesis of ATP form membrane localized
ATP synthase
n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

I
C
un|t
n
+

Sta|k
I
1
un|t
AD +
|
A1
The picture shows transport of H
+

through the ATP synthase.

What sort of transport would this be?

A. Passive diffusion
B. Facilitated diffusion
C. Active transport
D. Cant tell without more information
n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

I
C
un|t
n
+

Sta|k
I
1
un|t
AD +
|
A1
If the flow of protons were reversed,
from the direction shown, what sort
of transport would it be?

A. Passive diffusion
B. Facilitated diffusion
C. Active transport
D. Cant tell without more information
Membrane localized
ATP synthase
passively transports
H
+

down the
electrochemical
gradient.
Electrochemical gradient also called the Proton Motive Force
Captures energy that becomes available to phosphorylate ADP.
?
<
-#($10"#- 45
@9A 15$-.(1'
7#"3 9"
B<C B8C
?,;. D?
<
E F"G D?
<
E
Membrane fragments
showing the ATP
synthase stalk and F1
heads
n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

n
+

I
C
un|t
n
+

Sta|k
I
1
un|t
AD +
|
A1
Certain chemicals make the
membrane very permeable to
protons. When you add one of
these to bacteria or mitochondria,
protons pass freely in and out
and ATP synthesis stops. This
experiment supports which of the
alternatives as the source of the
energy needed to add P
i
to ADP?

A. The ATP synthase complex
B. The proton gradient
C. The movement of protons
D. ATP
2
222
2:
22
ETC is a series of
spontaneous redox
reactions
Last redox reaction:
O
2
is e
-
acceptor =
terminal electron
acceptor (TEA)
Reduced to H
2
O
Energy captured from the
decrease in free energy
over the series of reactions
is used to transport H
+


Bigger picture: a cells metabolic grid
Glycolysis and Krebs cycle
Metabolism
Catabolism
Iats form v|a dehydrat|on react|ons. Iats cons|st of g|ycero| ||nked by ester ||nkages to three fatty ac|ds.
G|ycero|
Iatty ac|d
Carbon atoms in fats are more highly reduced than in carbohydrates and
contain more high energy electrons. Fats can be oxidized and energy of
electrons captured to form ATP.
I|gure 9-29
Fats, fatty acids, amino acids, can also be oxidized
via the Krebs cycle (acetyl-CoA intermediate)
Anabolic reactions depend on molecules that are
intermediates in the catabolic reactions of glucose
oxidation
Week 12 - Biology 112

Short lecture followed by

activity


2
222
2:
22
ETC is a series of
spontaneous redox
reactions
Last redox reaction:
O
2
is e
-
acceptor =
terminal electron
acceptor (TEA)
Reduced to H
2
O
Energy captured from the
decrease in free energy
over the series of reactions
is used to transport H
+


Fermentation
Modes of catabolism
Respiration
Aerobic Anaerobic
Fermentation
Cell making ALL of its
ATP by substrate level
phosphorylation
Some bacteria use other
TEAs
eg. NO
3
-
, SO
4
-2

Cell making MOST of
its ATP by oxidative
phosphorylation
TEA = O
2
TEA = O
2
Fermentation
Eukaryotic cells
Some cells with mitochondria ferment when O
2
is in short
supply, e.g.:
Root cells of some plants when soil becomes waterlogged
Yeasts (eukaryotic microbes) in cultures with low dissolved O
2
Skeletal muscle cells during vigorous exercise
Brain cells of some fish in water with low dissolved O
2
Some cells must ferment because they are genetically
incapable of respiration, i.e. no genes for ETC
components. Ferment even in presence of O
2
.
Bacterial cells
Some cells ferment when O
2
(or other TEAs, anaerobic
respiration) are in short supply.
Some cells must ferment because they lack mitochondria
and thus have no ETC and no way of using O
2
as a TEA.
Eukaryotic cells (cont)
eg. red blood cells, some eukaryotic microbes.
Fermentation (of glucose)
NADH cannot be used to make ATP by oxidative
phosphorylation: either no ETC or no TEA
In the absence of O
2
as a TEA, NAD
+
cannot be
regenerated again to carry electrons - needs to be
alternative way to oxidize NADH
2

TEA
Regenerated
In fermentation NAD
+
is regenerated because NADH
donates e
-
to pyruvate or a derivative of pyruvate
Allows glycolysis to continue producing ATP via substrate-
level phosphorylation in the absence of a TEA e.g., O
2

Fermentation yields a fermentation by-product = waste product
Fermentations often named for their waste products
Skeletal muscle cells
Red blood cells
and
Some bacteria
Root cells of some
plants
Waste
Lactic acid fermentation
Waste
Waste
and
Some bacteria
Brain cells of
some fish
Root cells of
some plants
Ethanol fermentation
Comparing Glucose Respiration and Fermentation
Glucose respiration yields far more free energy and thus ATP
synthesized than fermentation ( ~ 10X more ATP)
Fundamental reason
Compared to fermentation, products of respiration are
more oxidized and much more stable than reactants.
Mostly due to enthalpy not entropy, i.e products much more
strongly bonded than reactants.
G =
H = - 669 kcal/mol
TS =
Glucose respiration (25
o
C)
Collection of
bonds in
products much
stronger than
collection of
bonds in
reactants
H =
100% -680 kcal/mol
-669 kcal/mol
-11 kcal/mol
98%
2%
Fermentation (example lactic acid fermentation, 25
o
C)
G =
TS =
H =
100%
H = -46 kcal/mol
Compared to
respiration,
collection of
bonds in products
not much stronger
than collection of
bonds bonded in
reactants
-47 kcal/mol
-46 kcal/mol
-1 kcal/mol 2%
98%
Now its time for an activity.