Enzymes Chapter 6

Enzymes
-Enzymes are mostly proteins , or catalytic RNAs.
Ribozymes (ribo nucleic acid en zyme s) are RNA molecules that have role in
RNA splicing in gene expression, similar to the action of protein enzymes.
- Highly specialized proteins catalyze biochemical rxn’s (high degree of specificity to
substrates)
- function at (mild conditions, temperature, neutral pH, aqueous environment)
- Activity depend on integrity of the native protein structure.
Denaturation / dissociation into subunits/ hydrolysis into a.a  lost activity  activity
dependent on structure ( primary, secondary, tertiary and quaternary.
- Mwt = 12,000 to million
- Measurement of enzyme activities in blood plasma and tissue samples diagnosing
Some enzymes require no chemical groups for activity other than
their amino acid residues.

Some require nonprotein

1) cofactors (metal ion)
2) coenzymes (organic)
-Some require both.
-Coenzyme / cofactor tightly and covalently bound= prosthetic group.
-Enzyme + cofactor and or coenzyme = holoenzyme
-Enzyme alone = apoprotein / apoenzyme
Cofactors for enzymes
Coenzymes
Coenzymes act as transient carriers of specific functional groups.

Most are derived from vitamins, organic nutrients required in small amounts in the
diet.

some enzyme proteins are modified covalently by phosphorylation, glycosylation,

and other processes.

Many of these alterations are involved in the regulation of enzyme activity

any of a class of yellow water-soluble nitrogenous pigments derived from
isoalloxazine and occurring in the form of nucleotides as coenzymes
of flavoproteins; especially :RIBOFLAVIN
Enzyme names:
1) Addition of suffix “ ase” to a name describing substrate / function
Urease  urea hydrolysis
DNA polymerase  polymerization of DNA

2) Named after the broad function before specific function known:

Pepsin = greek “ pepsis” = digestion
Lysozyme = lysis of bacterial cell wall

3) Some have 2 / more names .

Solution: System for naming and classification :6 classes, subclasses based on rxn catalyzed.

ATP + glucose  ADP + glucose -6 p

(ATP:glucose phosphotransferase)
Other name hexokinase

Class (transferase), subclass (phosphotransferase), phosphotransferase with hydroxyl as acceptor, glucose

as phosphoryl group acceptor.
Enzymes are classified according to the type of reaction they catalyze

6 classes based on rxn catalyzed.

Enzyme Classification

• As an example, the formal systematic name of the enzyme

catalyzing the reaction:
ATP + D-glucose ADP + D-glucose 6-phosphate
*** ATPglucose phosphotransferase: it catalyzes the transfer of a
phosphoryl group from ATP to glucose.
• All enzymes have formal EC numbers and names.
• Its Enzyme Commission number (E.C. number) is 2.7.1.1.
• The first number (2) denotes the class name (transferase);
• The second number (7), the subclass (phosphotransferase);
• The third number (1), a phosphotransferase with a hydroxyl group
as acceptor;
• and the fourth number (1), D-glucose as the phosphoryl group
acceptor.
How Enzymes Work?
 S P
S P

EE E E E

E+S ES EP E+P

E: enzyme / S: substrate / P: product

ES and EP are transient complexes of the enzyme with the substrate and with
the product
 Substrate binding to enzyme active site:
Chymotrypsin with S (red).

Active site: pocket of the E with

a.a residues with groups that
bind S

Substrate: molecule/ ligand

bound to active site and
acted on by the E.

Active site is lined with amino acid residues

with substituent groups that bind the substrate
and catalyze its chemical transformation
Enzyme Catalysis
• What is the function of the catalyst?
• to increase the rate of a reaction. The role of enzymes
is to accelerate the inter conversion of S (Substrate) and
P (product). Not effect on the both concentrations.
• Characteristics:
1. do not affect reaction equilibria.
2. The enzyme is not used up in the process, and the
equilibrium point is unaffected.
3. The reaction reaches equilibrium much faster when the
appropriate enzyme is present, because the rate of the
reaction is increased.
Decrease the time that needed for particular reaction
 Reaction coordinate diagram for a chemical rxn:
A description of energy changes during a rxn SP

Standard free energy

change ∆G° ( for standard set
conditions) pressure
of each gas 1 atm /
101.3 KPa, temp 25°C,
conc. of solutes 1M.

Biochemical systems
[H+] << 1M
P <<S
Biochemical standard -ve overall standard
Free-energy change
free energy change starting point In SP directions.
∆G`° at pH 7
rxn progress (e.g., bond breakage or formation)

∆G`° is the overall standard free-energy change in the direction S P.

To describe the free-energy changes for reactions, chemists
define a standard set of conditions (temperature 298 K; partial
pressure of each gas I atm, or 101.3 kPa; concentration of
each solute 1 M) and express the free-energy change for this
reacting system as ΔGo, the standard free energy change
 Energy barrier between S and P: required for Alignment of reacting groups, formation
of transient unstable charges, bond rearrangements
➢ For the reaction to occur:
must overcome this barrier and therefore must be raised to a higher
Parially breakdown, partially formed
energy level. The highest possible free energy
energy hill
➢ At the top of the energy hill is a point
called the transition state: where the bond is
starting to form and the geometry is changing. Activation energy
(bond breakage, bond formation, and charge A-B.C
development, ) have proceeded to the precise
point at which decay to either substrate or
product is equally likely. A+B-C

(The difference between the energy levels

of the ground state and the transition state
is the activation energy, G‡)
➢ The rate of a reaction reflects the activation energy
Higher activation energy  slower rxn

In cells, enzymes and other catalysts lower the activation energy

of a rxn enhance rxn rate.
accelerate interconversion of S and P

- E not used up in the rxn

Equilibrium constant Keq =
K = [P] / [S]
∆G`° = large and –ve 
favorable rxn.

the equilibrium constant

is directly related to the
overall standard free energy
change for the reaction
(Table 6–4). A large
negative value for G reflects
a favorable reaction
equilibrium—but as already
noted, this does not mean
the reaction will proceed at a
rapid rate
Reaction equilibria are linked to the standard free-energy
change for the reaction ∆G°

Reaction rates are linked to the activation energy, ∆G‡

Exergonic reaction will be spontaneous, negative ∆G°

Endergonic reaction will not be spontaneous and will need

energy to do, positive ∆G° https://www.youtube.com/watch?v=tPCOEUo6J8s
Sucrose C12H22O11 + O2  12CO2 + 11 H2O ∆G`° large and –ve
In vitro ( in container) : no rxn
In vivo : rxn + useful form of energy ATP
E overcomes the E barrier , important for control in metabolism.

Reaction intermediate: S ESEPP

Rxn with several steps:

Step/s with highest activation energy
= highest energy point in the diagram of SP interconversion = rate limiting
step.

E: enzyme / S: substrate / P: product

ES and EP are transient complexes of the enzyme with the substrate and with
the product
➢ The function of a catalyst is to increase the rate of a reaction

Any reaction may have several steps, involving the formation and
decay of transient chemical species called reaction intermediates.
- Enzymes are highly effective catalyst enhancing rxn rates by a factor of 105 -1017
Rate is described by determining how quickly the reactant are consumed or the product is
formed.
➢ Rate equation
Rate / velocity V = k[S] , k= rate constant, unit : reciprocal time: s-1
reflects probability of rxn in a given temp, pH, etc…
Order of reaction describes how the rate of reaction depend on conc.

➢ The rate of any reaction is determined by the concentration of the reactant (or
reactants) and by a rate constant (constant not depend on concentration).
S  P , 1st order rxn
If a first-order reaction has a rate constant k of 0.04 (mean that 4% of the
available S will be converted to P in 1 s)
A reaction with a rate constant of 2,000 s1 will be over in a small fraction of a
second
V = k[S]

S1 + S2  P , 2nd order rxn

If a reaction rate depends on the concentration of two different compounds,
or if the reaction is between two molecules of the same compound, the
reaction is second order and k is a second-order rate constant, with
units of M1s1.
V = k[S1] [S2] , k = M-1s-1
Now we turn from what enzymes do to how they do it?
Catalytic power and specificity of enzymes:

1) Transient covalent interactions bw S and catalytic site of E mainly functional

groups of some a.a, metal ion or coenzyme.
Rearrangement of bonds/ transfer of a functional group to E
➢ Covalent interactions lower the activation energy by providing an alternative,
lower-energy reaction path

2) Non covalent interactions bw E and S ( hydrophobic / ionic / H-bonds)

Formation of weak interaction in the ES complex is accompanied by release of a
small amount of free energy that provides a degree of stability to the interaction
• Each bond formation associated with energy release stabilize transition
state.
• Free energy released in forming bonds + interaction bw E and S
= binding energy = ∆GB

Binding energy is a major source of free energy used by

enzymes to lower the activation energies of reactions
➢How does an enzyme use binding energy to lower the activation energy
for a reaction?
Enzyme specificity :

1) Key and Lock model : ( misleading ) An enzyme completely

complementary to its substrate would be a very poor enzyme .
Emil Fischer 
E structurally complementary to S Fit in like key in lock
Does not explain how product released or the rate
Enzyme is dynamic not rigid

2) Induced fit :
Active site not 100% complementary to S but to transition states.
 An imaginary enzyme (stickase) designed to catalyze breakage of a metal stick

ES complex is more
stable and has less free
energy in the ground
state than substrate
alone. The result is an
increase in the activation
energy.
No change in E or
substrate shape

enzyme must be
complementary to the
reaction transition state
More acceptable model.
Involve parts of the stick far away from bent site stabilize + Changes in shape
away from water. Explain why E are large?
Weak Interactions between Enzyme and
Substrate Are Optimized in the Transition State
 Role of binding energy is catalysis:
activation energy for a rxn lowered by binding energy bw E and S weak
noncovalent interactions in transition state.

The summation of the

unfavorable (positive) activation
energy G‡ and the favorable
(negative) binding energy GB
results in a lower net activation
energy
Binding energy explains the need for large enzymes, and
account for specificity.

An enzyme must provide functional groups for ionic,

hydrogen-bond, and other interactions, and also must
precisely position these groups so that binding energy is
optimized in the transition state
Once a substrate is bound to an enzyme,
functional groups aid in the cleavage and formation of
bonds by a variety of mechanisms including:
1. acid-base catalysis
2. covalent catalysis
3. metal ion catalysis.
Several mechanisms of catalysis:
Acid- base catalysis: (general not specific –proton transfer involve molecules other than water)
In E active site several a.a serve as proton donors / acceptors (most common
catalysis type). Proton transfer provide rate enhancement of 102 – 105
Covalent catalysis:
Transient covalent bond formed bw S and E
A-B (H2O) A+B (hydrolysis of the bond)

Enzyme with nucleophile X: electron-rich atom

A-B + X:  A-X + B (H2O) A + X: +B
functional groups of some enzyme cofactors can serve as nucleophiles in the
formation of covalent bonds with substrates
Metal ion catalysis:
weak bonding interactions between metal and substrate

Ionic interactions bw E-bound metal + S

1) Orient S /or stabilize transition state.
2) Metals mediate redox rxns by reversible changes in metals oxidation state.

1/3 E require one or more metal ions for catalysis.

Most E employ more than one catalysis rxn
 Covalent and general acid-base catalysis:
1st step in rxn is acylation step. (cleavage of peptide bond)
Hydroxyl group of Ser195 is the nucleophile. (formation of a covalent linkage)
Catalytic triad in enzyme here

hydroxyl group of Ser195 is the nucleophile

His 57

Covalent linkage bw
E and part of S
the base is the side chain of His57
H+
electron-
deficient atom
(an
electrophile). Break peptide bond
OH in ser is not strong nucleophile so the H will be removed by His N atom, that
were oriented in the correct position by the attraction with Asp. i.e stabilize the
histidine so it can carry the protonation, and convert Ser to more nuleophile by
converting OH to alcoxide that attack the carbon of the carbonyl group . So break
down of peptide bond.

https://www.youtube.com/watch?v=OY1WsqlcUdo
Enzyme kinetics

Most enzymes have certain kinetic properties in common

Rate of rxn and the effect of [S]

Initial rate = initial velocity = V°

 Effect of [S] on the initial velocity of an E – catalyzed rxn:
Michaelis constant = Km = [S] where Vo is half maximal.
The substrate concentration at which V0 is half Vmax
Hyperbolic (all E except regulatory)

Plateau like V
Saturation
kinetics
At low [S]  Vo ↑ linearly steady state, [ES] constant
over time
with [S].
V0 increases by smaller and
At higher [S] Vo ↑ by smaller smaller amounts in response
to increases in [S]
amounts.
Pre-steady state, E free, [ES] builds up
micro seconds
E saturated with S
[S]  but effect on V is smaller.
 !Hyperbolic curve
Michaelis-Menten Equation

The important of measuring the V0 because at the beginning of a reaction the

changes in [S] are negligible, so [S] can be treated as a constant.

• The rate equation for a one-substrate enzyme-catalyzed reaction.

• It is a statement of the quantitative relationship between the initial velocity
V0, the maximum velocity Vmax, and the initial substrate concentration [S], all
related through the Michaelis constant Km.
• Km has units of concentration.
• The reaction quickly achieves a steady state in which [ES] (and the
concentrations of any other intermediates) remains approximately constant
over time.
• V0 Substrate concentration
• (mmol/min) (mM)
• —————————————
• 2 1
• 12 5
• 25 11
• 35 50
• 44 200
• 48 500
 Interpreting Vmax and KM

V0 = Vmax [S] Vmax = Vmax [S]

KM + [S] 2 KM + [S]

1 = [S]
2 KM + [S]

Solving for KM, we get: KM + [S] = 2[S], OR

KM = [S], when V0 =1 Vmax
2
Michaelis - Menten equation / rate equation:
Dependence of initial velocity Vo on [S]

The equation
Vo = Vmax [S]
Km +[S]

Vo = 1/2 Vmax
Vmax = Vmax [S]
2 Km + [S]

Km= [S]

Equation and Km provide little information about chemical nature/ steps of the rxn.
Importance of Km in enzyme reactions

• The Michaelis constant (Km) is a means of characterizing an enzyme's affinity for a

substrate.
• The Km in an enzymatic reaction is the substrate concentration at which the
reaction rate is half its maximum speed.
• A low Km value means: the enzyme has a high affinity for the substrate (as a
"little" substrate is enough to run the reaction at half its max speed).
• This is only true for reactions where substrate is limiting and the enzyme is NOT
allosteric.
Km indicator of the affinity of E to S.
Vary from E to another and from different S for the same E
E with low [S] in cell lower Km than E with abundant S (the lower Km the less substrate needed to form ES
complex
Michaelis-Menten Equation
Lineweaver–Burk Plot

V0 = Vmax [S]
KM + [S]
Double reciprocal plot or Lineweaver-Burk plot: (more convenient)
The Michaelis-Menten equation
Vo = Vmax [S]
Km+[S]

1 = Km+[S]
Vo Vmax [S]

Important in analyzing
E inhibition.

Advantage: precisely
determining Vmax
Kcat = most of the enzyme is in the EP form at saturation, and Vmax = k3[Et]. describes the limiting
rate of E – catalyzed rxn at saturation. kcat = Vmax/[Et]. The concentration of enzyme is necessary.
Kcat = turnover number = tell us the maximum number of S molecules  P in a unit time on a single
active site of E molecule when E saturated with S
EX:A single active site converts 40,000,000 substrates to product per sec
Taking the reciprocal of kcat(1/kcat: 1/40.000,000) give the time that is needed for
one substrate to convert to product.
Km: describes the amount of S needed for the E to obtain half of its
maximum rate of reaction.

km can be separated into two parts:

1) kd
The first step of catalysis kinetic is the binding between S and E
( rate determine step in the reaction).
• the better enzyme bind to substrate, the smaller kd = smaller kM

2) kcat
The second step of catalysis kinetic is the forming of P.
• The larger kcat is, the more favorable the reaction towards P
Kcat and Km for evaluation of kinetic efficiency of enzymes

Kcat = specificity constant

Km
kcat/KM = rate constant that measures catalytic efficiency.

➢ helpful in determining whether the rate is limited by the creation of

product or the amount of substrate in the environment.

if the rate of efficiency is:

•HIGH (kcat is much larger than KM) and the enzyme complex converts a
greater proportion of the substrate it binds into product. (substrate binds
more firmly (strong) to the enzyme, a consequence of relatively low KM ) or
a large turnover rate kcat.

•LOW (kcat is much smaller than KM ) and the complex converts a lesser
proportion of the substrate it binds into product
Enzymatic rxn with 2 or more S :

ATP + glucose hexokinase glucose-6-p + ADP

-Analyzed by M-M equation and M-M kinetics with Km for each S.

-Involve transfer of a functional group/ atom from one S to another.
 Bi-substrate Reaction
1)Sequential binding of S1 and S2 before catalysts:
A. Random substrate binding: either S1 or S2 can
bind first, then the other binds.
B. Ordered substrate binding: S1 must bind before S2.
2)Ping Pong reaction: first S1 give P1, then P1
released before S2 bind, then S2 give P2.
Common mechanisms for rxn with more than one S:
1) E and both S form a ternary complex.
In random binding substrates can bind in either order.
In ordered binding S1 must bind before S2 can bind productively.
2) Ping-pong or double-displacement mechanism:
E-S complex forms a P1 leaves the complex. Altered E forms a 2nd
complex with another S, and P2 leaves, regenerating E
S1 transfer a functional group to E to form covalently modified E’
which is transferred to S2 .
Enzyme inhibition:
inhibitors: agents that interfere with catalysis slowing / halting
enzymatic rxns.
1) Pharmaceutical agents:
e.g aspirin (acetylsalicylate) inhibits prostaglandins synthesis
(pain processes).
2) Discovery and define of metabolic pathway.

Inhibition is reversible / irreversible

Three types of reversible inhibition:
1) Competitive inhibitors:
Inhibitor resembles S and bind E  EI complex but no catalysis / P
αKm↑= “apparent”/ observed in I presence.
 Slope is larger (multiplied by a) Intercept does not change (Vmax is the same) KM is larger (multiplied
by a)

Solution:
Add more S
[S]>>[I]
 Competitive Inhibitor

 Most common
 Inhibitor competes with natural substrate for binding to
active site
 Inhibitor similar in structure to natural substrate and binds
active site of enzyme (reducing effective enzyme conc)
 Binds more strongly
 May or may not react
 If reacts, does so very slowly
 Gives info about active site through comparison of
structures
 Reversible Inhibition (competitive)

Vmax +Inh
1/v
-Inh
+inh
vo
-Inh
1/2 Vmax

1/Vmax

Km [Substrate] 1/[S]
-1/Km
α α
Inhibitor competes with substrates for binding to active site
Inhibitor is similar in structure to substrate
binds more strongly
reacts more slowly
Increasing [I] increases [EI] and reduces [E] that is available for substrate binding
Need to constantly keep [I] high for effective inhibition (cannot be metabolized away in body)
 Slope is larger (multiplied by a)
 Intercept does not change (Vmax is the same)
 KM is larger (multiplied by a)
Use of competitive inhibitor in Medical therapy:
Case:
methanol poisoning usually from contaminated alcohol beverages
Methanol in liver  alcohol dehydrogenase (EC 1.1.1.1)  formaldehyde  formic acid metabolic
acidosis
Symptoms:
Vomiting, abdominal pain, photophobia, tissue damage
Ingestion of 10ml blindness , 30ml  fatal (2 tablespoons deadly to a child).
Treatment:
Antidote‫ مضاد سمي‬to reverse the effect of the poison intravenous infusion of ethanol =
competitive inhibitor to alcohol dehydrogenase.
The dehydrogenase enzymes facilitate the interconversion between alcohols and aldehydes or ketones with the
. of nicotinamide adenine dinucleotide (NAD+ to NADH)
reduction
CH3OH + NAD+ CHO + NADH +H+ ethanol
CH3CH2OH + NAD+ → CH3CHO + NADH + H+ methanol
Uncompetitive inhibitors:
Bind at a separate site, but bind only to the ES complex ( after S)
α Km ↓ and Vmax ↓
Cyanide combines
with the prosthetic
group of cytochromo
oxidase and inhibits
the election
transport chain
1. Binds only to ES complex but not free enzyme

2. Binds at location other than active site

3. Does not look like substrate. Binding of inhibitor

distorts active site thus preventing substrate binding and
catalysis

4. Cannot be competed away by increasing conc of

substrate (Vmax is affected by [I])

Increasing [I] lowers Vmax and lowers Km.

  Increasing [I]
 Lowers Vmax (y-intercept Uncompetitive Inhibitor
increases)
 Lowers KM (x-intercept
decreases)
 Ratio of KM/Vmax is the
same (slope)
Vmax
-Inh
Vmax
(app)_
+inh
vo
1/2 Vmax
1/2 Vmax

‫ן‬Km Km [Substrate]
Mixed inhibitors:
Also bind to a site separate than the active site, but may bind to
either E or ES .
Inhibitor binds E or ES
 Increasing [I]

 Lowers Vmax (y-intercept

increases)
 Raises KM (x-intercept
increases)
 Ratio of KM/Vmax is not the
same (slope changes)

If the inhibitor may bind to the enzyme

whether or not the substrate has already
been bound, but has a higher affinity for
binding the enzyme in one state or the
other, it is called a mixed inhibitor.

Mixed-type inhibitors bind to both E and

ES, but their affinities for these two forms
of the enzyme are different.
 Reversible Inhibition (non-competitive)

A inhibitor binds the enzyme but not in its active site.

Rate of catalysis is affected.
Non-competitive inhibition is a type of enzyme inhibition where the inhibitor
reduces the activity of the enzyme and binds equally well to the enzyme whether or
not it has already bound the substrate. Non-competitive inhibitors have identical
affinities for E and ES.
i.e: non-competitive inhibition can occur with or without the substrate +Inh
present.

Vmax 1/v
-Inh
Vmax 1/Vmax -Inh
(app)_
+inh (app)
vo
1/2 Vmax
1/2 Vmax 1/Vmax
(app)

-1/Km 1/[S]
Km Km [Substrate]
(app)

Vmax is decreased proportional to inhibitor conc

It not afffect km
 1/v
+Inh

-Inh

-1/Km 1/[S]
Effects of reversible inhibitors on Km and Vmax

Inhibition Binding Km Vmax Km/Vma

x
Competitive E

Uncompetitive ES

Mixed ES, E

Noncompetitive E
Irreversible inhibition:
Binds irreversibly to E active site by forming a covalent bond/ very stable non
covalent interaction  destroy a functional group.

diisopropylflourophosphte (DIFP):
irreversibly inhibits chymotrypsin
Ser195 is the key active site residue in chymotrypsin.
Suicide inactivators / mechanism-based inactivators:
Relatively unreactive until binding to E active site.

Perform first few chemical steps of rxn not transformed to normal product.
• Inactivator is converted to a very reactive compound that combines irreversibly with the enzyme

Penicillin which inhibits transpeptidase from building bacterial cell walls.

Exemestane, a drug used in the treatment of breast cancer, is an inhibitor of
the aromatase enzyme.

Important in DRUG DESIGN:

Specific for a single E unreactive until within E active site. Advantage: few side effects
In biochemistry, suicide inhibition, also known as suicide
inactivation or mechanism-based inhibition, is an irreversible form of enzyme
inhibition that occurs when an enzyme binds a substrate analogue and forms an
irreversible complex with it through a covalent bond during the normal catalysis
reaction.

The inhibitor binds to the active site where it is modified by the enzyme to
produce a reactive group that reacts irreversibly to form a stable inhibitor-enzyme
complex.

This usually uses a prosthetic group or a coenzyme, forming electrophilic alpha

and beta unsaturated carbonyl compounds and imines
The pH activity profiles of two enzymes:
Optimum pH is close to the pH of the environment in which the
enzyme is normally found.
Optimum pH maximum activity.

Gastric juice pH = 1 - 2 pH of hepatocyte cytosol = 7.2

Regulatory Enzymes:
- One / more E in a metabolic pathway have great effect on the rate of
the overall rxn.
- Activity of allosteric E ( increase /decrease) regulated by reversible
binding of a specific modulator to a regulatory site other than active site.
Effect of modulator = positive / negative.
Modulators maybe the S itself (Homotropic modulator) / other
metabolites.
Kinetic behavior of allosteric E reflects cooperative interactions among E
subunits ( similar to O2 – hemoglobin).
Regulatory Enzymes:
Allosteric E regulated by reversible noncovalent binding of modulators
/ effectors.
Other E regulated by reversible covalent modification.
- All are (Multisubunit proteins).

Structural allosteric have 1 or more allosteric sites each with

a site specific to modulator.
Larger and more complex e.g. asparate transcarbomylase.
Regulatory enzyme aspartate transcarbamoylase (nucleotide synthesis):
12 polypeptide chains
2 catalytic clusters. Each with 3 polypeptide catalytic chains (blue + purple).
3 regulatory clusters. Each with 2 polypeptide regulatory chains (red + yellow).
Regulatory clusters form the points of a triangle surrounding catalytic subunits.
Subunit interactions in an allosteric E, interactions with inhibitors and
activators:

S binding site and M sites

on different subunits.
Catalytic regulatory
Binding of M to R induce
conformational change in C
active and capable of
binding S with higher affinity.

When M dissociates.
Enzyme inactive/ less
active.
In most multienzyme systems, the first enzyme
of the sequence is a regulatory enzyme

Feedback inhibition =
- The regulatory E in a metabolic pathway is
inhibited by the end product of the pathway.
- Build up of end product slows entire pathway

Heterotropic allosteric inhibition =

Noncovalent + reversible.
Pathway inhibited allostericaly by L-isoleucine.
But not by any of the 4 intermediate.
 Kinetics of allosteric enzymes:
Differ than M-M kinetics. Sigmoid saturation curve not hyperbolic.
Reflects cooperative interaction bw subunits.
reaction catalyzed by an allosteric enzyme
Small changes in [S]
Large effect on activity
K0.5 not Km

The sigmoid curve of

a homotropic enzyme
 Sigmoid curve for allosteric enzymes:
An activator cause the curve to become  more hyperbolic.
K0.5 decrease, no change in Vmax and large change in activity.
An inhibitor cause the curve to become  more sigmoid

activator
Resulting in an increased
reaction velocity at a fixed
inhibitor
substrate concentration
(V0 is higher for any value
of [S]

Ex: competitive inhibitor

A less common type of modulation, in which Vmax is
altered and K0.5 is nearly constant
Covalent modifications (reversible) of regulatory enzymes by a functional group
What is the role of serine and histidine at the active site
of serine proteases?

A) strong; weak
B) weak; strong
C) acid-base; covalent
D) covalent; acid-base
E) anionic; ionic