Reginald H.

Garrett
Charles M. Grisham

Chapter 15
Enzyme Regulation

Outline
What factors influence enzymatic activity ?
What are the general features of allosteric regulation ?
Can allosteric regulation be explained by
conformational changes in proteins ?
What kinds of covalent modification regulate the
activity of enzymes ?
Is the activity of some enzymes controlled by both
allosteric regulation and covalent modification ?
Special focus: is there an example in nature that
exemplifies the relationship between quaternary
structure and the emergence of allosteric properties ?
(hemoglobin and myoglobin paradigms of protein
structure and function).

15.1 What Factors Influence Enzymatic

Activity?
The availability of substrates and cofactors usually
determines how fast the reaction goes. As product
accumulates, the apparent rate of the enzymatic
reaction will decrease due to lack of substrate.
Enzyme activity can be regulated through covalent
modification.
Enzyme activity can be regulated allosterically.
Zymogens, isozymes, and modulator proteins may
play a role.
Genetic regulation of enzyme synthesis and decay
determines the amount of enzyme present at any
moment.

Levels of Enzyme Regulation

1. Enzyme level: The enzyme may be activated or
inhibited by either noncovalent or covalent
interactions. This is the most rapid control
system.
2. Hormonal level: A hormone is secreted and
carries a message to the cell and in turn an
enzyme is activated or inhibited. The speed of
this control system is intermediate.
3. Gene level: A message is sent to the nucleus
either express or repress a gene. This
determines the amount of enzyme produced
and is the slowest control measure.

Modes of regulation

Zymogens or proenzymes.
Enzyme cascades.
Isozymes.
Allosterism
Covalent modification.
Second messengers.
Combined effects

15.1 What Factors Influence Enzymatic

Activity?

Zymogens or proenzymes are

inactive precursors of enzymes.
Typically, proteolytic cleavage
produces the active enzyme.

Figure 15.2
Proinsulin is an 86-residue
precursor to insulin

Figure 15.3 The proteolytic activation of

chymotrypsinogen

Proteolytic Enzymes of the Digestive Tract

The Cascade of Activation Steps Leading to

Blood Clotting
Figure 15.4
The intrinsic and
extrinsic pathways
converge at factor X,
and the final common
pathway involves the
activation of thrombin
and its conversion of
fibrinogen into fibrin,
which aggregates into
ordered filamentous
arrays that become
crosslinked to form
the clot.

Isozymes Are Enzymes With Slightly

Different Subunits

Figure 15.5 The

isozymes of lactate
dehydrogenase (LDH).

15.2 What Are the General Features of

Allosteric Regulation?
Action at "another site"
Allosteric enzymes have quaternary structure and
regulation is at a site other than the active site.
Enzymes situated at key steps in metabolic
pathways are modulated by allosteric effectors.
The effectors are usually produced elsewhere in
the pathway (heterotropic). S or P = homotropic.
Effectors may be feed-forward activators or
feedback inhibitors.
Kinetics are sigmoid ("S-shaped"), see MM plot.

Kinetics of
allosteric enzymes
Generally these dont obey MichaelisMenten kinetics
Homotropic positive effectors produce
sigmoidal (S-shaped) kinetics curves
rather than hyperbolae
This reflects the fact that the binding of
the first substrate accelerates binding of
second and later ones

15.2 What Are the General Features of

Allosteric Regulation?

Figure 15.6 Sigmoid v versus [S] plot.

The dotted line represents the hyperbolic plot
characteristic of normal Michaelis-Menten kinetics.

15.2 What Are the General Features of

Allosteric Regulation?
Effects
A positive effector activates the enzyme (an
activator).
A negative effector inhibits the enzyme (an
inhibitor).
Positive cooperativity increases substrate binding
in an adjacent subunit.
Negative cooperativity decreases substrate
binding in an adjacent subunit.

15.3 Allosteric Regulation and

Conformational Changes in Subunits.
Monod, Wyman, Changeux (MWC) Model:
allosteric proteins can exist in two states: R
(relaxed) and T (taut or tight).
In this two-state model, all the subunits of an
oligomer must be in the same state (they all
change together) and is therefore termed the
concerted model.
T state predominates in the absence of
substrate S.
S binds much tighter to R than to T.

The Concerted Model for Allosteric

Regulation (MWC)

Figure 15.7 Allosteric effects:

A and I binding to R and T, respectively.

The Concerted Model for Allosteric

Regulation

Figure 15.7
Allosteric effects:
A and I binding
to R and T,
respectively.

The Concerted Model for Allosteric

Regulation

Figure 15.7 Allosteric effects:

A and I binding to R and T, respectively.

More about the MWC model

Positive cooperativity is achieved because S
binding increases the population of R, which
increases the sites available to S.
In the MWC ligands such as S are positive
homotropic effectors.
Molecules that influence the binding of
something other than themselves are
heterotropic effectors.
The MWC model does not explain negative
cooperativity.

The Sequential Model for Allosteric

Regulation (KNF)
An alternative model proposed by Koshland,
Nemethy, and Filmer (the KNF model) relies on
the idea that ligand binding triggers a
conformation change in a protein.
In this one-state model, ligand-induced
conformation changes in one subunit may lead to
conformation changes in adjacent subunits.
The KNF model explains how ligand-induced
conformation changes could cause subunits to
adopt conformations with little affinity for the
ligand i.e., negative cooperativity.
The KNF model is termed the sequential model.

The Sequential Model for Allosteric

Regulation
Figure 15.8
The Koshland-Nemethy-Filmer
sequential model for allosteric
behavior. (a) S binding can, by
induced fit, cause a conformation
change in the subunit to which it
binds. (b) If subunit interactions
are tightly coupled, binding of S to
one subunit may cause the other
subunit to assume a conformation
having a greater or lesser affinity
for S. That is, the ligand-induced
conformational change in one
subunit can affect the adjoining
subunit.

The Sequential Model for Allosteric

Regulation
Figure 15.8
The KoshlandNemethy-Filmer
model.
Theoretical curves
for the binding of a
ligand to a protein
having four identical
subunits, each with
one binding site for
the ligand.

Allosteric Models

MWC: Two state concerted

KNF: One state sequential

S

S
S

15.4 Covalent Modification Regulate the

Activity of Enzymes
Enzyme activity can be regulated through reversible
phosphorylation.
This is the most prominent form of covalent
modification in cellular regulation.
Phosphorylation is accomplished by protein kinases.
Each protein kinase targets specific proteins for
phosphorylation.
Phosphoprotremoving phosphoryl groups from
proteins.ein phosphatases catalyze the reverse
reaction
Kinases and phosphatases themselves are targets of
regulation.

15.4 Covalent Modification Regulate the

Activity of Enzymes
Enzymes that catalyze phosphate transfer
Kinase:
An enzyme that transfers phosphate to ADP or AMP
or from ATP.
Phosphatase or phosphoprotein phosphatase:
An enzyme that hydrolyzes a phosphate off of a
substrate.
Phosphorylase:
An enzyme that adds a phosphate to substrate but
does not use ATP to do so.

15.4 Covalent Modification Regulate the

Activity of Enzymes

Figure 15.1 Enzyme regulation by reversible covalent

modification. Depending on the enzyme,
phosphorylation may activate or inactivate its catalytic
function.

15.4 Covalent Modification Regulate the

Activity of Enzymes
Protein kinases phosphorylate Ser, Thr, and Tyr
residues in target proteins.
Kinases typically recognize specific amino acid
sequences in their targets.
In spite of this specificity, all kinases share a
common catalytic mechanism based on a conserved
core kinase domain of about 260 residues, Fig. 15.9.

15.4 Covalent Modification Regulate the

Activity of Enzymes

MAPK Kinase

Human ERK5 (MAPK7) protein domains. NES1 and NES2, bipartite

nuclear exportation signal; PB1-BD, PB1 (Phox and Bem domain 1)
binding domain; Kinase Domain, catalytic kinase domain; TEY,
sequence motif containing ERK5 regulatory phosphorylation
residues; PR-1 and PR-2, proline rich domains; Transcriptional transactivation, transcriptional activity domain. The ERK5 N-terminus domain
resembles the typical MAPK catalytic domain and includes the MAPKconserved TXY activation sequence (T218EY220) in the activation loop. The
activation of ERK5 occurs via interaction with and dual
phosphorylation in its TEY motif by MKK5 (Mody et al., 2003). MKK5
mediated ERK5 activation leads to ERK5 autophosphorylation in its
unique C-terminal domain (Morimoto et al., 2007).

15.4 Covalent Modification Regulate the

Activity of Enzymes
Figure 15.9
Protein kinase A is shown
complexed with a
pseudosubstrate peptide
(orange).
This complex also includes
ATP (red) and two Mn2+ ions
(yellow) bound at the active
site.

Cyclic AMP-dependent protein kinase

Figure 15.10 cyclic AMP-dependent protein kinase, also

known as protein kinase A (PKA), is a 150- to 170-kD R2C2
tetramer in mammalian cells.
The two R (regulatory) subunits bind cAMP; cAMP binding
releases the R subunits from the two C (catalytic) subunits.
C subunits are enzymatically active as monomers.

Other Covalent Modification of Protein

Several hundred different chemical modifications
of proteins have been discovered.
Only a few of these are used to achieve metabolic
regulation through reversible conversion of an
enzyme between active and inactive forms.
A few are summarized in Table 15.3.
Three of the modifications in Table 15.3 require
nucleoside triphosphates (ATP, UTP) that are
related to cellular energy status.

Other Covalent Modification of Protein

15.5 Enzymes Controlled by Both Allosteric

and Covalent Regulation?
Glycogen phosphorylase (GP) is an example of the
many enzymes that are regulated both by allosteric
controls and by covalent modification.
GP uses Pi to attack glucose at an (1-4) linkage
on the nonreducing ends of glycogen.
This converts glycogen into readily usable fuel in
the form of glucose-1-phosphate.
This is a phosphorolysis reaction.
Muscle GP is a dimer of identical subunits, each
with PLP covalently linked.
There is an allosteric effector site at the subunit
interface.

Glycogen Phosphorylase

Figure 15.11 The glycogen phosphorylase reaction converts

glycogen into readily usable fuel in the form of glucose-1-P.
The enzyme cannot cleave an (1-6) branchpoint.

Phosphoglucomutase

Figure 15.12 The phosphoglucomutase reaction converts

glucose-1-P into the glycolytic substrate, glucose-6-P.

The structure of glycogen phosphorylase

Glycogen phosphorylase is a dimer of identical
subunits each having 842 residues.
Each subunit contains an active site (at the center of
the subunit) and an allosteric effector site near the
subunit interface.
A regulatory phosphorylation site is located at Ser14
on each subunit.
A glycogen-binding site exerts regulatory control.
Each subunit contributes a tower helix (residues
262 to 278) to the subunit-subunit interface.
In the dimer, the tower helices extend from their
respective subunits and pack against each other.

The structure of glycogen phosphorylase

Figure 15.13
The structure of
glycogen
phosphorylase.
One monomer of
the homodimer.

Glycogen Phosphorylase Activity is

Regulated Allosterically
Muscle glycogen phosphorylase shows
cooperativity in substrate binding.
ATP and glucose-6-P are allosteric inhibitors of
glycogen phosphorylase.
AMP is an allosteric activator of glycogen
phosphorylase.
When ATP and glucose-6-P are abundant,
glycogen breakdown is inhibited.
When cellular energy reserves are low (i.e., high
[AMP] and low [ATP] and [G-6-P]) glycogen
catabolism is stimulated.

Glycogen Phosphorylase Activity is

Regulated Allosterically

Figure 15.14 v versus S curves for glycogen phosphorylase.

(a) The response to the concentration of the substrate
phosphate (Pi).
(b) ATP is a feedback inhibitor.
(c) AMP is a positive effector. It binds at the same site as ATP.

Glycogen phosphorylase conforms to the

MWC model
The active form of the enzyme is designated the
R state.
The inactive form of the enzyme is denoted the T
state.
AMP promotes the conversion to the active state.
ATP, glucose-6-P, and caffeine favor conversion to
the inactive T state.
A significant conformation change occurs at the
subunit interface between the T and R state.
This conformational change at the interface is
linked to a structural change at the active site that
affects catalysis.

Glycogen Phosphorylase is Controlled

Both Allosterically and Covalently

Figure 15.15
The mechanism of covalent
modification and allosteric
regulation of glycogen
phosphorylase.

Favored

Favored

A Conformation Change Regulates Activity

of Glycogen Phosphorylase
Figure 15.16
The major conformational
change that occurs in the
N-terminal residues upon
phosphorylation of Ser14.
Ser14 is shown in red.
N-terminal conformation of
phosphorylated enzyme
(phosphorylase a): yellow.
N-terminal conformation of
unphosphorylated enzyme
(phosphorylase b): cyan.

Regulation of GP by Covalent Modification

In 1956, Edwin Krebs and Edmond Fischer
showed that a converting enzyme could
convert phosphorylase b to phosphorylase a.
Three years later, Krebs and Fischer show that
this conversion involves covalent
phosphorylation.
The enzyme is phosphorylase kinase and this
phosphorylation is mediated by an enzyme
cascade (Figure 15.17).

Glycogen phosphoryase is activated by a

cascade of reactions

Figure 15.17 The hormone-activated enzymatic cascade

that leads to activation of glycogen phosphorylase.

The Adenylyl Cyclase Reaction

Figure 15.18 The adenylyl cyclase reaction. The reaction is

driven forward by subsequent hydrolysis of pyrophosphate by
the enzyme inorganic pyrophosphatase.

The Adenylyl Cyclase Reaction

(Shown here are
the products of the
reaction. ATP, the
reactant, is shown
on the previous
slide.)

Figure 15.18 The adenylyl cyclase reaction. The reaction is

driven forward by subsequent hydrolysis of pyrophosphate by
the enzyme inorganic pyrophosphatase.

cAMP is a Second Messenger

Cyclic AMP is the intracellular agent of
extracellular hormones - thus a second
messenger.
Hormone binding stimulates a GTP-binding
protein (G protein), releasing G(GTP).
Binding of G(GTP) stimulates adenylyl cyclase to
make cAMP.

cAMP is a Second Messenger

Figure 15.19 Hormone binding to its receptor leads via
G- protein activation to cAMP synthesis. Adenylyl
cyclase and the hormone receptor are integral membrane
proteins; G and G are membrane-anchored proteins.

Modes of regulation

Zymogens or proenzymes.
Enzyme cascades.
Isozymes.
Allosterism
Covalent modification.
Second messengers.
Combined effects

Hemoglobin
A classic example of allostery
Hemoglobin is an oxygen transport protein
and myoglobin is an O2 storage protein.
Look at the oxygen binding curves for
hemoglobin and myoglobin.
Myoglobin is monomeric; hemoglobin is
tetrameric, 22 .
Mb: 153 aa, 17,200 MW.
Hb: two chains of 141 residues, 2 chains
of 146 residues.

Figure 15.20 O2-binding curves for

hemoglobin and myoglobin

Myoglobin P50 = 2.8 torr and Hemoglobin P50 = 26 torr.

The structure of myoglobin is similar to that

of the Hb monomer
Figure 15.21
The myoglobin and
hemoglobin structures.
Each is a protein with a
heme (aromatic).
Myoglobin is monomeric.
Hemoglobin is tetrameric.

Mb and Hb use porphryins to bind Fe 2+

Figure 15.22
Heme is formed when protoporphyrin IX binds Fe2+

Fe2+ is coordinated by His F8

Iron interacts with six ligands in Hb and Mb.
Four of these are the N atoms of the porphyrin.
A fifth ligand is donated by the imidazole side
chain of amino acid residue His F8.
(This residue is on the sixth or F helix, and it is
the 8th residue in the helix, thus the name.).
When Mb or Hb bind oxygen, the O2 molecule
adds to the heme iron as the sixth ligand.
The O2 molecule is tilted relative to a
perpendicular to the heme plane.

Fe2+ is coordinated by His F8

Figure 15.23
The six liganding
positions of an
iron atom in Hb
and Mb.

Myoglobin Structure
Mb is a monomeric heme protein.
Mb polypeptide "cradles" the heme group.
Fe in Mb is Fe2+ - ferrous iron - the form that
binds oxygen.
Oxidation of Fe yields 3+ charge - ferric iron.
Mb with Fe3+ is called metmyoglobin and does
not bind oxygen.

O2 Binding Alters Mb Conformation

In deoxymyoglobin, the ferrous ion actually lies
0.055 nm above the plane of the heme.
When oxygen binds to Fe in heme of Mb, the
heme Fe is drawn toward the plane of the
porphyrin ring.
With oxygen bound, the Fe2+ atom is only 0.026
nM above the plane.
For Mb, this small change has little consequence.
But a similar change in Hb initiates a series of
conformational changes that are transmitted to
adjacent subunits.

Hb Has an 22 Tetrameric Structure

Figure 15.24
An dimer of Hb,
with packing
contacts indicated in
blue.
The sliding contacts
made with the other
dimer are shown in
yellow. The changes
in these sliding
contacts are shown
in Figure 15.25.

Cooperative Binding of Oxygen Influences

Hemoglobin Function
Mb, an oxygen storage protein, has a greater
affinity for oxygen at all oxygen pressures.
Hb is different it must bind oxygen in lungs and
release it in capillaries.
Hb becomes saturated with O2 in the lungs, where
the partial pressure of O2 is about 100 torr.
In capillaries, pO2 is about 30 torr, and oxygen is
released from Hb.
The binding of O2 to Hb is cooperative binding of
oxygen to the first subunit makes binding to the
other subunits more favorable.

O2 binding curves of Mb and Hb

The oxygen binding curve of

Mb resembles an
enzyme:substrate saturation
curve.

An Alternative O2 Binding Curve for Hb

Oxygen saturation
curve for Hb in the
form of Y versus pO2
assuming n = 4 and
P50 = 26 torr.
Y is the fractional
saturation of Hb.
4

[ pO2 ]
Y
4
[ pO2 ] K

An Alternative O2 Binding Curve for Hb

A comparison of the
experimentally
observed O2 curve for
Hb yielding a value for
n of 2.8,
the hypothetical curve
if n = 4,
and the curve if n = 1
(non-interacting O2binding sites).

The Conformation Change

The secret of Mb and Hb

Oxygen binding changes the Mb conformation.

Without oxygen bound, Fe2+ is out of heme plane.
Oxygen binding pulls the Fe2+ into the heme plane.
Fe2+ pulls its His F8 ligand along with it.
The F helix moves when oxygen binds.
Total movement of Fe2+ is 0.029 nm i.e., 0.29 .
This change means little to Mb, but lots to Hb!

Oxygen Binding by Hb Induces a

Quaternary Structure Change
When deoxy-Hb crystals are exposed to oxygen,
they shatter. This is evidence of a large-scale
structural change.
One alpha-beta pair moves relative to the other
by 15 degrees upon oxygen binding.
This massive change is induced by movement of
Fe by 0.039 nm when oxygen binds.

Oxygen binding to Hb results in a 15

rotation of one pair relative to the other

Figure 15.25 Subunit motion in hemoglobin when the

molecule goes from the (a) deoxy form to the (b) oxy form.

Fe2+ Movement by Less Than 0.04 nm

Induces the Conformation Change in Hb
In deoxy-Hb, the iron atom lies out of the heme
plane by about 0.06 nm.
Upon O2 binding, the Fe2+ atom moves about 0.039
nm closer to the plane of the heme.
As if the O2 is drawing the heme iron into the plane.
This may seem like a trivial change, but its
biological consequences are far-reaching.
As Fe2+ moves, it drags His F8 and the F helix with
it.
This change is transmitted to the subunit interfaces,
where conformation changes lead to the rupture of
salt bridges.

Fe2+ Movement by Less Than 0.04 nm

Induces the Conformation Change in Hb
Figure 15.26
Changes in the
position of the heme
iron atom upon
oxygenation lead to
conformational
changes in the
hemoglobin
molecule.

Salt bridges that stabilize deoxy-Hb are

broken in oxy-Hb
Figure 15.27
Salt bridges between different
subunits in human deoxy-Hb.
These noncovalent, electrostatic
interactions are disrupted upon
oxygenation.
(a) The salt bridges and H-bonds
involving interactions between Nterminal and C-terminal residues
in the -chains.
(b) The salt bridges and H bonds
involving C-terminal residues of chains

The Physiological Significance of the

Hb:O2 Interaction
Hb must be able to bind oxygen in the lungs.
Hb must be able to release oxygen in capillaries.
If Hb behaved like Mb, very little oxygen would
be released in capillaries - see Figure 15.20!
The sigmoid, cooperative oxygen binding curve
of Hb makes its physiological actions possible!
Hb exhibits properties of both the MWC and
KNF models.
In the T state only the subunits can bond O2
and in the R state all subunits can bind O2.

H+ Promotes Dissociation of Oxygen from

Hemoglobin
Binding of O2 to Hb is affected by several agents,
including H+, CO2, 2,3-bisphosphoglycerate and
chloride ions.
The effect of H+ is particularly important. This is
the Bohr effect.
Deoxy-Hb has a higher affinity for H+ than oxy-Hb.
Thus, as pH decreases, dissociation of O2 from
hemoglobin is enhanced.
Ignoring the stoichiometry of O2 and H+, we can
write:

H+ Promotes Dissociation of Oxygen from

Hemoglobin

Figure 15.28
The oxygen saturation curves for myoglobin and for
hemoglobin at five different pH values: 7.6, 7.4,7.2, 7.0, 6.8.

The Antagonism of O2 Binding by H+ is

Termed the Bohr Effect
The effect of H+ on O2 binding was discovered by
Christian Bohr (the father of Neils Bohr, the atomic
physicist).
Binding of protons diminishes oxygen binding.
Binding of oxygen diminishes proton binding.
Important physiological significance.

HHbO2 <==> HbO2 + H+

HHb <==> Hb + H+

pKa = 6.6
pKa = 8.2

Figure 15.20 O2-binding curves for

hemoglobin and myoglobin

Myoglobin P50 = 2.8 torr and Hemoglobin P50 = 26 torr.

CO2 Also Promotes the Dissociation of O2

from Hemoglobin
Carbon dioxide diminishes oxygen binding
1. Hydration of CO2 in tissues and extremities leads
to proton production:

These protons are taken up by Hb as oxygen

dissociates.
The reverse occurs in the lungs.
2. CO2 also binds covalently to the N-terminus. And
fovors the T state.

Summary of the Physiological Effects of H+

and CO2 on O2 Binding by Hemoglobin
At the tissue-capillary interface, CO2 hydration
and glycolysis produce extra H+, promoting
additional dissociation of O2 where it is needed
most.

At the lung-artery interface, bicarbonate

dehydration (required for CO2 exhalation)
consumes extra H+, promoting O2 binding.

2,3-Bisphosphoglycerate
An Allosteric Effector of Hemoglobin
In the absence of 2,3-BPG, oxygen binding to
Hb follows a rectangular hyperbola!
The sigmoid binding curve is only observed in
the presence of 2,3-BPG.
Since 2,3-BPG binds at a site distant from the
Fe where oxygen binds, it is called an allosteric
effector.

BPG Binding to Hb Has Important

Physiological Significance

Figure 15.30 The structure, in ionic form of BPG or 2,3bisphosphoglycerate, an important allosteric effector of Hb.

BPG Binding to Hb Has Important

Physiological Significance
The "inside" story......
Where does 2,3-BPG bind ?
"Inside.
in the central cavity of the tetramer.
What is special about 2,3-BPG ?
Negative charges interact with 2 Lys, 4 His,
2 N-termini.
Fetal Hb - lower affinity for 2,3-BPG, higher
affinity for oxygen, so it can get oxygen from
mother.

BPG Binding to Hb Has Important

Physiological Significance
Figure 15.31
The ionic binding of
BPG to the two subunits of Hb. BPG
lies at the center of the
cavity between the two
-subunits.
The resulting
conformational change
precludes O2 binding.

CO2 Also Promotes the Dissociation of O2

from Hemoglobin
Figure 15.29
Oxygen binding curves of blood
and of hemoglobin in the
absence and presence of CO2
and BPG.

Fetal Hemoglobin Has a Higher Affinity for

O2 Because it has a Lower Affinity for BPG
The fetus depends on its mother for O2, but its
circulatory system is entirely independent.
Gas exchange takes place across the placenta.
Fetal Hb differs from adult Hb with -chains in
place of -chains and thus a 22 structure.
As a result, fetal Hb has a higher affinity for O2.
Why does fetal Hb bind O2 more tightly ?
Fetal -chains have Ser instead of His at position
143 and thus lack two of the positive charges in
the BPG binding cavity
BPG binds less tightly and Hb F thus looks more
like Mb in its O2 binding behavior.

Fetal Hemoglobin Has a Higher Affinity for

O2 Because it has a Lower Affinity for BPG

Figure 15.32 Comparison of the oxygen saturation curves

of Hb A and Hb F under similar conditions of pH and [BPG].

Sickle-Cell Anemia is a Molecular Disease

Sickle-cell anemia patients have abnormallyshaped red blood cells.
The erythrocytes are crescent-shaped instead of
disc-shaped.
The sickle cells pass less freely through the
capillaries, impairing circulation and causing tissue
damage.
A single amino acid substitution in the -chains of
Hb causes sickle-cell anemia.
Glu at position 6 of the -chains is replaced by Val.
As a result, Hb S molecules aggregate into long,
chainlike polymeric structures.

Hemoglobin and Nitric Oxide

Nitric oxide (NO) is a simple gaseous molecule
that acts as a neurotransmitter and as a second
messenger in signal transduction (see Chapter
32).
NO is a high-affinity ligand for Hb, binding to the
heme iron 10,000 times more tightly than O 2.
So why is NO not bound instantaneously to Hb,
preventing its physiological effects ?
NO reacts with the SH of Cys93, forming an Snitroso derivative:

Hemoglobin and Nitric Oxide

The S-nitroso group is in equilibrium with other Snitroso compounds formed by reaction of nitric
oxide with small-molecule thiols such as free Cys
or glutathione:

These small-molecule thiols transfer NO from

erythrocytes to endothelial receptors, where it
exerts its physiological effects.

End Chapter 15
Enzyme Regulation