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ATP: Adenosine Triphosphate

Professor B. Z. Chowdhry
School of Science
Room 202, Grenville Building
Tel: 0208-331-8208
E-mail: b.z.chowdhry@gre.ac.uk

ATP has the following structure:

ATP consists of: a base (adenine), a sugar (ribose) and a


phosphate chain.

ATP possesses 2 phosphoanhydride bonds.


AND
ATP possesses 1 phosphoester bond

The following statements relating to ATP are


incorrect:
a) that it can trap and store energy
b) that it is the universal energy currency
c) it is a high energy molecule
d) that it can give up its stored energy in order
to drive reactions which otherwise would be
energetically favourable
e) that it is an unstable molecule.

The following statements are also


untrue:
ATP is hydrolysed to ADP and Pi
phosphate transfer potentials account for the
function of ATP
the coupling of oxidation of substrates i.e.
catabolism to anabolism is energetic not
stoichiometric

What about:
High energy phosphate compounds
Low energy phosphate compounds ?
Concept of high energy and low energy
phosphates not useful!

ATP is claimed to have special roles in energy coupling and


phosphate transfer. The free energy of hydrolysis of phosphate
from ATP is intermediate among the examples listed in Table
1 (next slide). ATP thus act as a phosphate donor (true), and
ATP can be synthesized by transfer of phosphate from other
compounds, such as phosphoenolpyruvate (PEP).
BUT Go OF PHOSPHATE HYDROLYSIS IS IRRELEVANT

Table 1. Standard free energies of hydrolysis


Compound

Go' of phosphate hydrolysis


(kJ/mol)

Phosphoenolpyruvate (PEP)

61.9

Phosphocreatine

43.1

Pyrophosphate

33.5

ATP (to ADP)

30.5

Glucose-6-phosphate

13.8

Glycerol-3-phosphate

9.2

THE FACTS

1) The rate of ATP turnover in cells is seconds to


minutes.
2) ATP cannot be stored.
3) ATP is water soluble.
4) Under physiological conditions ATP is stable.
5) ATP in cells, is complexed to Mg2+ ions i.e.
ATP-Mg2+ complex is structurally complex (can
be recognized easily and specifically without
ambiguity by enzymes).
6) ATP is involved in reactions which are
irreversible.

8) Reactions involving ATP in vivo are never


close to equilibrium.

9) Cellular reactions are under kinetic control


not thermodynamic control.

10) Free energy (Go) values apply to


reactions, not reactants (or products).

11) ATP is a very important phosphorylating


agent for both small molecular weight
molecules (e.g. glucose) and proteins.
12) Reactions which involve ATP are usually
catalysed by kinase enzymes e.g., hexokinase
and phosphofructokinase reactions in glycolysis.
13) ATP occurs in all eukaryotic and most
procaryotic cells; it is central to many cellular
functions.

ATP is produced via three main


mechanisms:
(a) substrate level phosphorylation (glycolysis
and the TCA cycle)
(b) oxidative phosphorylation ( occurs in the
mitochondria of eukaryotic cells; the majority of
ATP is produced via this mechanism)
(c) the adenylate kinase reaction (mainly in
muscle cells as an emergency supply of ATP).

ATP can be used in a number of synthetic


reactions which can be generalised as follows:
XOH + YOH + nATP + (n-1)H2O

XOY + nADP + H3PO4 (1)

If n = 1 then equation (1) can be split into sets of


hypothetical reactions:
XOH + YOH
XOY + H 2O
ATP + H2O
ADP + H 3PO4
OR
XOH + ATP
XOP + ADP
XOP + YOH
XOY + H 3PO4

(2)
(3)
(4)
(5)

Which of these sets of reactions describes the in


vivo synthesis of XOY?

(i) the amount of XOY formed in the


presence of enzymes catalysing (2) and
(3) depends only on the equilibrium
constant for reaction (2) and is
independent of the presence of ATP.
(ii) reaction (3) does not achieve anything
and is very unlikely to occur in vivo
because ATP is hardly ever hydrolysed!

Reactions (4) and (5) are plausible because the


amount of XOY which is formed will depend on
the concentrations of XOH, YOH and ATP.
Reactions (4) and (5) are chemically i.e.
stoichiometrically linked.
Values of free energies (Go) for the above
reactions are irrelevant (cellular systems are
under kinetic not thermodynamic control).

ATP is being continuously synthesized and used.


Cannot be stored
Metabolic half life of ATP seconds to minutes
Brain cells only a few seconds supply of ATP

Living systems are not at equilibrium


Non-equilibrium thermodynamics
Oscillating thermodynamics
(Complicated ??)

The real role of ATP has nothing to do with energetics


and
is a gross misuse as well as
misunderstanding of thermodynamic rules
..
This does not detract from the fact that in cells
ATP fulfils vital functions which need to be
understood if progress is going to be made in
understanding cellular function

The primary causes of the present situation in


which the energetics of biological processes
are discussed in misleading terms lies in two
major areas:
(i)free energy (Go) values apply to reactions,
not reactants (or products).
(ii)the inability to distinguish between kinetic
and thermodynamic control.

All reactions in vivo are enzyme catalysed and


biological systems operate by complicated
sequences of integrated reactions in which
very few, if any, of the individual steps are at
equilibrium.
Living systems are under kinetic not
thermodynamic control.
Furthermore it is likely that reactions involving
ATP in vivo are never close to equilibrium.

The intermediate has to have the following


properties:
(i)
be stable under physiological conditions
and water soluble.
(ii)
have some degree of structural
complexity:
Recognition
Specificity

ATP should be involved in reactions which


are irreversible
Large fluctuations in the levels of ATP
concentration would disrupt integrated
metabolic pathways
The directions in which metabolic reactions
proceed will be under kinetic control. This is
achieved by changes in concentrations of
reactant(s) or product(s) or by the effects of
reactants, products and positive or negative
effectors.

There are many examples of coupled reactions involving


ATP in vivo, these include:
(i)

the synthesis of nucleotides and polysaccharides,

(ii)

the chemical activation of amino acids as esters of


a specific tRNA prior to their incorporation into
proteins,

(iii)
the synthesis of acetylcoenzyme A in bacterial
cells.

If ATP is to function in its many roles its


concentration in cells has to be maintained at
adequate levels.
It must not be destroyed by direct hydrolysis.
It is apparent that the central role of the
nucleotide in metabolism is due as much to its
kinetic versatility as to its thermodynamic
properties.
Reminder:
It is a fundamental thermodynamic property of
chemical reactions that the equilibrium constant
of a reaction reflects only the equilibrium
concentrations of reactants and products and is
completely independent of the pathway which
the reaction follows.

There are many examples of coupled reactions


involving ATP in vivo, these include the:
(i)synthesis of nucleotides and polysaccharides.
(ii) chemical activation of amino acids as esters of
a specific tRNA prior to their incorporation into
proteins.
(iii)synthesis of acetylcoenzyme A in bacterial
cells.
It is worth examining reaction (iii) in more
detail.

Consider the following reactions:


(1) CH3CO2- + ATP4-

CH3COOPO32- + ADP3-

(2) CH3COOPO32- + HSCoA

CH3COSCoA + HOPO32-

Reactions (1) and (2) sum to the following reaction:


(3) CH3CO2- + ATP4- + HSCoA

CH3COSCoA + ADP3- + HPO42-

The equilibrium constants for reactions (1) and (2) are:


8 x10-3 and 74, respectively.
[Note that the forward and backward reactions would be
catalysed, in vivo, by different enzymes!]

For the overall reaction (3) the equilibrium constant


(Keq) is equal to: 0.59 (i.e. 8 x10-3 x 74).
For direct esterification Keq = 6.6 x 10-5.
Question
Why is the Keq for reaction (3) much larger than that for
the direct condensation reaction?

Answer
In any set of coupled reactions the elements of water (not a
molecule of water as such) are removed from the reactants of
the condensation and incorporated into phosphate.
If we make allowances for ionisation an OH group is removed
from acetic acid (CH3CO2-) and liberated as part of ADP whilst
H+ is removed from coenzyme A and liberated as part of
phosphoric acid.
The formation of the condensation product is therefore
facilitated because water is being removed and liberated in the
form of a species which has mole fractions less than unity!

Vitamin
source

Major metabolic role

Mechanistic role

Coenzyme
Adenosine triphosphate

Transfer of phosphoryl
or
nucleotidyl groups

Cosubstrate

Metabolite Coenzymes:
Synthesized from common metabolites
Include: e.g. ATP
Methionine + ATP

adenosylmethionine + PPPi
(S-AM)

S-AM is required for:


the conversion of norepinephrine to epinephrine
HO

Norepinephrine

CH2

CH2

+
NH3

HO

HO

Epinephrine
HO

CH2

CH2

+
NH2

CH3

Topic 2
Protein phosphorylation

Protein phosphorylation catalyzed by protein


kinases plays a critical role in many cellular
processes e.g. cellular signalling

Protein phosphorylation

During protein phosphorylation by ATP


phosphate moieties are usually transferred to
the serine, threonine or tyrosine (sometimes
arginine) amino acid residues of proteins from
ATP molecules by protein kinases

Reversible protein phosphorylation acts as a


biological regulatory mechanism
It is thought that perhaps 30 % of the proteins
encoded by the human genome contain
covalently bound phosphate, and abnormal
phosphorylation is now recognized as a cause
or consequence of many human diseases

Protein + ATP
Active

E1

Protein-P +ADP
Inactive

OR
Protein + ATP
Inactive

E2

Protein-P +ADP
Active

Signalling networks
Can be very complex
E.g. protein A phosphorylates protein B
Protein B phosphorylates C
BUT
A can also phosphorylate C directly
B can phosphorylate D, which may in turn phosphorylate A

Phosphorylation of sugars: part of their catabolism


allows cells to accumulate sugars

30 % of all proteins in cells may undergo phosphorylation


Human genome : 2 % of genes may be kinase genes

Disregulated kinase activity: cause of


disease e.g. cancer
Kinases regulate many aspects of cell
growth, movement and death
Attempts are being made to develop drugs
to inhibit specific kinases to treat several
diseases e.g., the drugs:
Gleevec (imatinib)
Iressa (gefitinib)