respiration

Cyclic metabolic pathways

Oxidation of glucose
1

Students will understand how energy is

transformed from chemical or solar
energy into the universal power source
of cells, ATP.

Focus: Photosynthesis
Cyclic Metabolic pathways
Overview of Respiration
Feedback control of electron-stripping reactions of respiration
2

Cyclic metabolic pathways

initial
substrate
intermediate
#1

enz.1

interintermediate
mediate
#2
#3
enz.2

enz.3

inter- enz.4
mediate
#4

intermediate
#1

product

Each turn of the cycle produces a product and reconstitutes

3
the initial starting molecule

Cyclic metabolic pathways

initial
substrate
intermediate
#1

enz.1

enz.2

enz.4
intermediate
#4

intermediate
#2

enz.3

intermediate
#3

Otherwise, they follow

all the rules of a
metabolic pathway

Fixing a pathway if an
enzyme is broken
becomes more
problematic...

product

Each turn of the cycle produces a product and reconstitutes

4
the initial starting molecule

Cyclic metabolic pathways

What will happen if
enzyme 2 is blocked?

enz.1

A. nothing- it works fine

B
enz.2

enz.4

enz.3

product

B. product can be made

as long as there is A
C. product can be made if
supplied with C
D. product can be made if
supplied with D
E. everything halts
immediately
5

Cyclic metabolic pathways

What will happen if
enzyme 4 is blocked?

enz.1

A. nothing- it works fine

B
enz.2

enz.4

enz.3

product

B. product can be made

as long as there is A
C. product can be made if
supplied with C
D. product can be made if
supplied with E
6

Cyclic metabolic pathways

What do I need to know?

initial
substrate
intermediate
#1

enz.1

intermediate
#2
enz.2

enz.4
intermediate
#4

initial substrates

enz.3

product

intermediate
#3

products
what leaves
what is reconstituted

highlighted enzymes & how

they are regulated
(different for each pathway)

...and what happens if you

interfere with an enzyme
7

The Calvin cycle is a cyclic metabolic pathway

inputs:
CO2, ATP, NADPH

initial substrates:
CO2, RuBP

cycle product:
G3P

reconstituted product:
RuBP

important enzymes:
Rubisco

enzyme regulation
(none discussed...

instead understand how and why some

plants use C4 and CAM photosynthesis)
8

The Citric Acid cycle is another cyclic pathway

inputs:
Acetyl CoA, NAD+, FADH+
Initial substrates:
NAD
+

Acetyl CoA

NADH

oxaloacetate
NAD
+

NADH

Cycle products:
CO2 (waste)

ADP
ATP
NADH

FADH
+
NAD
+

FADH2

ATP (or GTP)

3 NADH, 1 FAHD2
Reconstituted product:
oxaloacetate
important enzymes:
Step 1: citrate synthase

(well discuss this one later)

Step 39& 4: dehydrogenases

G3P is freshly-fixed carbon it has many

uses in the cell
reconstitute
ATP

glycolysis

oly
s is

G3P

Make starch
Make fats

gly
c

Make glucose

g
gluco

enesi

Make cellulose

e
os
t
n
Pe

Make NAD+
and NADP+

Make nucleotides

os
h
p

e
at
h
p

hw
at

ay

Make vitamin B1

Make some
amino acids

Make ATP/ADP
make DNA, RNA

make proteins

build other organic

molecules

10

10

Overview of respiration and

photosynthesis
PHOTOSYNTHESIS

GLUCOSE:

RESPIRATION

CO2 + H2O

temporary storage

CO2 + H2O

Catabolic
pathways

Anabolic
pathways

ATP

Organic molecules of the cell

11

11

BIG PICTURE: Respiration

moved into mitochondrial matrix- must cross 2 membranes

in cytoplasm

in mitochondrial matrix

occurs across
christae membrane
ATP made into matrix

Figure 9.2

12

12

The first three steps of RESPIRATION are about

stripping high-energy electrons from glucose

Figure 9.2

13

13

The first three steps of RESPIRATION are about

stripping high-energy electrons from glucose

Note that these first three steps also make a little ATP themselves.
We call this substrate-level phosphorylation.
This does not involve ATP synthase or the electron transport chain.
NOT enough ATP to support cells for long.
14
Figure 9.2

The high-energy electrons are taken to the

electron transport chains in mitochondria, where
they power proton pumps.

The proton gradient is used to create ATP.

We call this chemiosmosis using proton motive force.
Its also called oxidative phosphorylation.
Figure 9.2

15

15

The first three steps of RESPIRATION are about

stripping high-energy electrons from glucose

NAH+ and FADH+

The empty (oxidized) carriers return to the first three steps to collect more electrons.
Its essential that you have empty electron carriers for the
first three steps to work!
Figure 9.2

16

The first three steps of RESPIRATION are about

stripping high-energy electrons from glucose
ADP

ADP

ADP

Its also essential that you have ADP available to make ATP

Figure 9.2

17

17

The first three steps are

standard metabolic pathways
standard

questions apply!
...only one additional layer of regulation:
phosphorylation

of enzymes
allosteric feedback inhibition

18

Glycolysis is a standard metabolic pathway

G3P
Challenge: name one enzyme that catalyzes an exergonic reaction.
(How can you tell?)
figure 9.5....note that enzyme names are in table 9.1

19

Glycolysis is a standard metabolic pathway

G3P

Are these ATP molecules made through ATP synthase?

figure 9.5....note that enzyme names are in table 9.1

20

Cells control metabolic pathways with

positive and negative feedback.
energy pathways in general
In energy-producing processes, its not the intermediates that feed back to
regulate enzymes, its the cells level of energy
Low energy indicators:

High energy indicators:

High ADP, AMP

High ATP

High NAD+

High NADH

High FADH+

High FADH2

Generally stimulate

Generally repress

(positive feedback)

(negative feedback)
21

Cells control metabolic pathways with

built-in feedback: Glycolysis
Enzyme 3:
Phosphofructokinase

If the cell has enough ATP at the moment, ATP is the regulatory
G3P
molecule that
allostericly inhibits phosphofructokinase.
3

P
ATP
Note: This is not the same as turning it off via phosphorylation
22
figure 9.5....note that enzyme names are in table 9.1

Cells control metabolic pathways with

built-in feedback: Glycolysis
Enzyme 3:
Phosphofructokinase

Inhibition is temporary and reversible

G3P

When energy levels drop, ATP will come off and be used in other reactions.
Enzyme turns on again.
Challenge: why choose this particular enzyme to regulate glycolysis?
(see p. 160)
23
figure 9.5....note that enzyme names are in table 9.1

Cells control metabolic pathways with

built-in feedback: Pyruvate processing

Enzyme: Pyruvate dehydrogenase

complex
Inhibited by: phosphorylation

high NADH and high Acetyl CoA levels

INCREASE pyruvate dehydrogenase
phosphorylation
high CoA levels and high NAD+ levels
DECREASE pyruvate dehydrogenase
phosphorylation
24

Cells control metabolic pathways with

built-in feedback: Citric Acid cycle

We will discuss this more

next week

25

Please review the questions on the

following slides before recitation

26

Rubisco is a surprising enzyme

Eight identical Large subunits
Eight identical Small subunits
Eight active sites

Three reactions / second

(So: 2 seconds to produce one
molecule of glucose per enzyme)

Its SLOW
Is Rubisco encoded by a gene? If so, how many?
27

27

Rubisco is a surprising enzyme

Oxygen competes with CO2 at
the active sites.
Photorespiration is the
addition of O2 to RuBP it
takes energy (ATP) to undo
this reaction.

Its INEFFICIENT
Where is the O2 coming from....?

28

28

Oxygen competes with carbon

dioxide for rubiscos active sites.
That makes oxygen:
A.
B.
C.
D.
E.

an allosteric inhibitor
a noncompetitive inhibitor
a competitive inhibitor
a cofactor
a coenzyme
29

How have plants adapted to cut

down on photorespiration, to
maximize carbon fixation?
Add more enzyme
B. Add more CO2
A.

C.

Add more O2

D.

Decrease O2
30

30

How have plants adapted to cut

down on photorespiration, to
maximize carbon fixation?
STRATEGY ONE: add more enzyme
Make a TON of Rubisco.
Rubisco accounts for 50% of the protein in
some leaves
31

31

How might we increase CO2

(substrate) concentrations relative
to A.O2prevent
(inhibitor)
concentrations?
all gas exchange by closing stomata
B.
C.
D.
E.

actively pump O2 out of cells, or pump CO2 in

move rubisco reactions away from light
reactions
store CO2, release it all at once to
concentrate it
perform rubisco reactions at night when light
reactions are off and O2 production is low
32

32

How have plants adapted to cut

down on photorespiration, to
maximize carbon fixation?

STRATEGY TWO: move rubisco reactions

away from light reactions
CO2 temporarily attached to
PEP carboxylase
Moved to another cell
released into Calvin cycle

33

33

How have plants adapted to cut

down on photorespiration, to
maximize carbon fixation?

STRATEGY THREE: store CO2, release it

all at once to concentrate it
CO2 temporarily stored as
organic acid at night when stomata
are open
CO2 released into Calvin cycle
during the day when stomata are
closed.
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34

How have plants adapted to cut

down on photorespiration, to
maximize carbon fixation?
STRATEGY FOUR: shrug and deal with it
...then find a way to use the products in
other processes like cell-cell signalling
and development
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35

You should be able to

For each step of respiration, tell me:

Where in the cell it occurs
What the major regulatory enzymes are
The input and output of each pathway (including how many molecules of ATP,
NADH, FADH2 are made)
The overall equation of respiration and how glucose (C6H12O6)is converted into
CO2 and H2O

Looking at the metabolic pathways and electron transport chains,

predict which steps are endergonic/exergonic.
Predict the effects of artificially blocking any enzyme in these pathways
(Reminder: there are other metabolic pathways that feed into this one)

Compare/contrast the two processes that make ATP in respiration

Substrate level phosphorylation (glycolysis, citric acid cycle)
Oxidative phosphorylation (ATP synthase, proton gradient)

...and photosynthesis

photophosphorylation (ATP synthase, proton gradient)

36

You should be able to

Explain the difference between inhibition of an enzyme

via :

phosphorylation
competitive inhibition
allosteric regulation
...and how in each of these situations we could manipulate either
enzymes, substrates, or regulatory molecules to increase
reaction rates

Explain how the cell regulates the energy-producing

metabolic pathways (examples from class(

Which enzymes of which processes are regulated this way

Which molecules are the inhibitors and how do they inhibit?
Whether the inhibition is reversible, and how
What it means to the cell to have low ATP or high NAD+ levels
37