Glycolysis – Part II

Chapter 17

Outline
• stage 2 of glycolysis
- glyceraldehyde-3-phosphate converted to pyruvate
• regulating glycolysis
• anaerobic metabolism of pyruvate
• energy yields of glycolysis
the main objective of glycolysis is to convert six-carbon glucose into
two three-carbon molecules of PYRUVATE

there are five types of reactions used in glycolysis:

Phosphorylation – phosphate group is added to substrate by a KINASE
- there is a subset of this reaction type called PHOSPHATE TRANSFER
when a phosphate group is taken from one substrate and added
to another

Isomerization – the rearrangement of some of the groups on the

substrate molecule

Cleavage – substrate is cut in half

Redox

Dehydration – one oxygen and two hydrogens are removed from

substrate and released as a water molecule
• where did we leave off…?

• on Monday, we were able to take glucose

and turn it into two molecules of
glyceraldehyde-3-phosphate (GAP)

• now, everything we’re going to do will

be done in duplicate!

• one glucose molecule yielded two

GAP molecules
- those two GAP molecules
will be processed in
parallel (as seen →)

• today, we’ll make some ATP

- the PAYOFF PHASE
• so, let’s get to it
 Step 6
• in this step, we find our first bona fide REDOX reaction

• also, this step is really a two-for-one

- along with the redox reaction is a phosphorylation
- we will discuss these two aspects of the reaction separately
(to keep it as simple as possible)
- but, keep in mind that they are occurring simultaneously

• oxidations result in substrate gaining oxygen

- let’s look at that…
 Step 6
• what’s reduced….?

• it’s a simple reduction of NAD+ to NADH + H+

- our co-enzyme doing its thing to allow redox to proceed

• this redox component of Step 6 is exergonic

- it has a ΔG of -43.1 kJ/mol

• but remember, the redox component is only half of this reaction

 Step 6
• the phosphate is being added as a DEHYDRATION reaction

• this phosphate group is not being donated by ATP (first time ATP is NOT
our phosphate donor)
- instead, it is simply free phosphate (Pi) floating in the cell

• water is removed, leaving phosphate hungry to make a bond

- it makes that bond with OXIDIZED glyceraldehyde-3-phosphate
the result of the
• this reaction is endergonic (ΔG=49.3 kJ/mol) oxidation reaction
 Step 6
• let’s take another look at the two individual components
• the overall reaction is
very slightly
REDOX endergonic
ΔG=6.2 kJ/mol

• but remember, from

this point on in
glycolysis we
are doing
everything in…
PHOSPHORYLATION PARALLEL or
DUPLICATE

• so the ΔG really equals

12.4 kJ/mol
 Step 6
• the enzyme responsible for converting glyceraldehyde-3-phosphate
into 1,3-bisphosphoglycerate is glyceraldehyde-3-phosphate
dehydrogenase
- this enzyme belongs to a class of
enzymes called NADH-LINKED
DEHYDROGENASES
- enzymes that are linked to
and use NAD+ to steal
hydrogens (dehydrogenate) from
substrate
- remind me again why this is bis…
Step 6
 Step 6
• there’s one very important thing to know
about this molecule…

• it REALLY wants to give up the phosphate from

carbon 1
- and I mean REALLY….

• we call this a HIGH PHOSPHATE TRANSFER

POTENTIAL – meaning it has a high
potential for transferring its phosphate

• in fact, 1,3-bisphosphate wants to give that phosphate up MORE than

ATP wants to give its phosphate up

• in other words, 1,3-bisphosphate has a higher phosphate transfer

potential than ATP and so can be used to phosphorylate ADP
 Step 6
• some rich snot just bought himself a super-HD mega-screen TV

• he’s got no use anymore for his old, but

fine, normal big-screen TV

• he’s got a HIGH TV TRANSFER POTENTIAL

- higher than mine
- what he doesn’t want, I’ll take

• he gives up his normal TV and I grab it up

- with all my money going to daycare,
I’ll take any TV I can find

• 1,3-bisphosphate has a higher phosphate

transfer potential than ATP
- the phosphate that 1,3-bisphosphate
gives up will be grabbed by the less finicky, more hungry ADP
 Step 7
• now, finally, we’re going to make ATP (by phosphorylating ADP)

• this reaction is catalyzed by PHOSPHOGLYCERATE KINASE

• a phosphate group is transferred from 1,3-bisphosphate directly to ADP

- this leaves 3-phosphoglycerate and ATP

• the reason this reaction works – and is energetically favorable – is

1,3-bisphosphate wants to lose that phosphate group so badly
 Step 7
• the energy contained in the bond between the phosphate group and

carbon 1 is higher than the energy required to put that phosphate

group on ADP
- the universe still makes an energy profit
- (-49.3 kJ/mol) vs. (30.5 kJ/mol)
- so, this reaction will proceed
• this is SUBSTRATE LEVEL PHOSPHORYLATION (for obvious reasons…)
 Step 7

• please remember,
everything we do
today in GLYCOLYSIS
we do in duplicate
- because of the
isomerization
of DHAP into
GAP
- so we just made two
molecules of ATP

- and, since we
burned two ATP
on Monday in
Stage 1, we also
just BROKE
EVEN
 Step 8
• in this step, we simply move the phosphate group of
3-phosphoglycerate from carbon 3 to carbon 2, making…
- 2-phosphoglycerate

• this is a prep step for Step 9

• it is catalyzed by PHOSPHOGLYCEROMUTASE
 Step 8
• for our purposes, we can consider this an ISOMERIZATION
- because it is an “intra-molecular phosphate transfer” reaction,
hardcore chemists would disagree
- that’s why the enzyme does not get the name ‘… isomerase’
- but, as far as we’re concerned, this is a rearrangement of groups
that were already present on the molecule
- and, therefore, an isomerization
Step 8
 Step 9
• our second DEHYDRATION reaction

• 2-phosphoglycerate loses one oxygen and two hydrogens

- these atoms come together and leave as water

• what’s left is phosphenolpyruvate (PEP) – an extremely important

molecule
• the enzyme catalyzing this reaction is ENOLASE
and, looky here…
- we’re very close!
Step 9
 Step 10
• our last step of GLYCOLYSIS – we will make pyruvate in step 10

• the phosphate group on PEP has a very high transfer potential

- much higher than the phosphate transfer potential of ATP
- (-61.9 kJ/mol) vs. (30.5 kJ/mol)

• what can we expect to happen if we mix PEP and ADP…?

• and that is exactly what happens…

- PEP loses it’s phosphate to ADP making ATP and leaving pyruvate
 Step 10
• the double bond on carbon 2 shifts - carbon 3 takes on a third proton

• this is another example of SUBSTRATE LEVEL PHOSPHORYLATION and

PHOSPHATE TRANSFER

• and remember, this is done in duplicate, so we’ve just made two

molecules of ATP and pyruvate
- we had broken even before with respect to ATP, so these two
are profit
- PYRUVATE KINASE catalyzes this reaction, and is inhibited by ATP
 Step 10

• and that’s the end of

glycolysis
 Controlling Glycolysis
• this pathway is much too costly and much too important to leave
unregulated
- so, how do we control the rate of glycolysis?

• if a cell does not need ATP, glycolysis is pointless and wasteful

• three reactions of glycolysis are control points

- places where regulation is achieved

• we discussed two of them on Monday

- step 1: the phosphorylation of glucose to glucose-6-phosphate
is regulated by feedback inhibition (hexokinase)
- step 3 and step 10 are both catalyzed by allosteric enzymes
- phosphofructokinase and pyruvate kinase, respectively
- these enzymes are inhibited by ATP itself
• this allows communication between the cell and glycolysis about ATP
Controlling Glycolysis
 Anaerobic Metabolism of Pyruvate
• we’re most concerned with aerobic respiration and how pyruvate feeds
into the citric acid cycle
we’ll begin discussing that next week

• but let’s take a little time now to discuss how pyruvate is used for
energy without oxygen
- anaerobic metabolism of pyruvate
• anaerobically, pyruvate can be reduced to lactate (NADH is oxidized)
 Anaerobic Metabolism of Pyruvate
• this reaction is catalyzed by LACTATE DEHYDROGENASE (LDH)

• each reaction has a ΔG of -25.1 kJ/mol and we do it twice for every

glucose molecule used – so we’re getting ~50 kJ/mol out of this

• however, lactate is a “dead end” in our muscle cells

- it must be sent to the liver where enzymes exist to recycle it
back into pyruvate (and even glucose, if necessary)
- GLUCONEOGENESIS (coming up next lecture)

• the formation of lactate also helps to

recycle reduced NADH back to
its oxidized form (NAD+)
- without this, anaerobic cells
would run out of NAD+
and glycolysis would stop
 Anaerobic Metabolism of Pyruvate
• as we all know, some organisms can also make alcohol anaerobically
- this requires two reactions

• first, pyruvate is DECARBOXYLATED (a reaction type where substrate

loses a carbon dioxide molecule – one carbon and two oxygens)

• this reaction is catalyzed by

PYRUVATE DECARBOXYLASE
and leaves acetaldehyde

• it is this carbon dioxide that gives

beer its bubbles

• second, acetaldehyde is reduced to

ethanol while NADH is oxidized to NAD+
- this also recycles NADH for the cell
 The Energy Yield of Glycolysis
• although the point of this pathway is not to make energy, but simply
to make pyruvate for the citric acid cycle
- we do make some energy here

• we profit two molecules of ATP

- and release 74 kJ/mol of energy into the universe (exergonic)
- not bad…

• if we didn’t capture some of that released energy in ATP, it would all

be lost as dissipated heat
- even the 74 kJ/mol that is generated is released mostly as heat
keeping us warm-blooded animals warm…

• we will see another interesting example of heat being generated by

metabolism for the sole purpose of warmth later
 Summary
• we completed glycolysis
• we oxidized and phosphorylated GAP
into 1,3-bisphosphoglycerate;
transferred a phosphate from BPG
to ADP making two molecules of
ATP; isomerized 3-PG into 2-PG;
dehydrated 2-PG into PEP and
had PEP transfer its phosphate to
ADP making two more ATPs
- thus giving us our 2 ATP
profit and PYRUVATE

• we also discussed how pyruvate is

used anaerobically
- and briefly discussed the
energy yields of glycolysis