ENZYMES

Lesson 14-2
 Proteins that catalyze
chemical reactions

Enzymes
Characteristics
Presence of enormous Regulation of catalytic
catalytic power High specificity in reactions activity by the cell
•Speeds up chemical reactions by catalyzed and the •Reaction rate and the amount of
lowering activation energies and formation of any product are
allow reactions to achieve substances involved controlled by regulating enzyme
equilibrium more rapidly action
An Energy Diagram for a Reaction When Uncatalyzed and Catalyzed
Enzyme Specificity
• Absolute specificity: Enzyme acts only on one substance
• Relative specificity: Enzyme acts on structurally related substances
• Stereochemical specificity: Enzyme has the ability to distinguish
between stereoisomers
Exercise
• What is the primary function of enzymes?
1. Catalysis
2. Movement
3. Storage
4. Structure
Exercise
• A catalyst will reduce the energy
associated with the _____
1. products
2. reactants
3. transition state
4. More than one answer is correct
Classifying and Naming Enzymes
• Earliest enzymes were given names with an -in ending to indicate
their protein composition
• Examples - Pepsin, trypsin, and chymotrypsin
• Enzyme Commission (EC) system
• Enzymes are grouped based on the type of reaction catalyzed
• Enzyme name specifies the substrate (substance acted on), functional group
acted upon, and type of reaction catalyzed
• Names end in -ase
The EC Classification of Enzymes
EXAMPLE OF THE EC
SYSTEM OF ENZYME
NOMENCLATURE

Hydrolysis of urea
Common Names for Enzymes
• Shorter than EC names
• Formed by adding -ase to the name of the substrate or to a
combination of the substrate name and type of reaction
• Examples
• Urea amidohydrolase - Common name is urease
Substrate: urea
Common name: urea + ase = urease
• Alcohol dehydrogenase - Name is derived from both the substrate name and
the type of reaction
Substrate: alcohol ( ethyl alcohol )
Reaction type: dehydrogenation (removal of hydrogen)
Common name: alcohol dehydrogenation + ase = alcohol dehydrogenase
Exercise
• Which of the types of enzymes is
responsible for catalyzing the joining of
two molecules?
1. Isomerases
2. Ligases
3. Lyases
4. Transferases
Enzyme Cofactors
• Nonprotein molecules or ions required by an enzyme for catalytic
activity
• Can either be coenzymes or inorganic ions
• Coenzyme: Organic cofactor that is often derived from vitamins
• Apoenzyme: Catalytically inactive protein formed by the removal of the
cofactor from an active enzyme
Apoenzyme + cofactor → active enzyme

• Inorganic ions are metal ions, such as Mg2+, Zn2+, and Fe2+
Vitamins and Their Coenzyme Forms
Exercise
• What type of reaction does N-methyltransferase catalyze?
1. Oxidation-reduction
2. Transfer of a functional group
3. Addition to a double bond
4. Isomerization
 • An organic cofactor is
called a(n) _____
1. active site
Exercise 2. apoenzyme
3. coenzyme
4. substrate
 • If an enzyme requires a Zn2+ ion in order to be an
active enzyme, the protein portion of the
enzyme is called a(n) _____

Exercise 1. apoenzyme
2. coenzyme
3. holoenzyme
4. isoenzyme
 All enzymes have an active site

Active site: Location on the enzyme where a substrate is bound and catalysis occurs

Mechanism of
Enzyme
Action Binding of the substrate may occur through
hydrophobic attraction, hydrogen bonding, and/or
ionic bonding
Enzyme–substance (ES)
Chemical transformation of After conversion, the
complex is formed when a
the substrate that occurs in product is released from the
substrate and enzyme bond,
the active site is aided by active site and the enzyme is
resulting in conversion of
enzyme functional groups ready to catalyze again
substrate (S) to product (P)
Mechanism of Enzyme Action - Example

• General reaction

• Specific example
Theories on
Enzyme
Mechanism
• Lock-and-key theory
• States that the substrate has a
shape that exactly fits the active
site
• Limitation
• Implies that enzyme
conformations are fixed or
rigid but research proves
otherwise
Theories on Enzyme
Mechanism (continued)
• Induced-fit theory
• Modification of the lock-and-key
theory
• States that the conformation of the
active site changes to accommodate
an incoming substrate
• Active site has a shape that
becomes complimentary to the
substrate only after the substrate
is bound
Exercise
• Which of the following statements could be made based ONLY on the
equation below?

1. The enzyme is a zymogen

2. The enzyme activity is regulated by feedback inhibition
3. The enzyme has stereochemical specificity in this reaction
4. The enzyme can be reused after this reaction sequence is complete
Exercise
• What is the substrate for histamine N-methyltransferase?
1. Histamine
2. N-methyl
3. Transfer
4. ase
Exercise
• A rigid _____ is associated with the
_____ mechanism of enzyme action
1. enzyme; induced-fit
2. enzyme; lock-and-key
3. substrate; induced-fit
4. substrate; lock-and-key
 Substances that decrease enzyme
activity
Characteristic function of many
poisons and medicines is to inhibit
one or more enzymes and decrease
the rate of catalytic reaction
Reasons to understand enzyme
inhibition

Enzyme Some substances found in cells

inhibit enzyme-catalyzed reactions
and provide a means for internal
regulation of cellular metabolism

Inhibitors Irreversible inhibitors

Classified into:

Reversible inhibitors
 • Covalently bond with a specific functional group
of the enzyme and render it inactive
• Include:
• Poisons
• Toxic metals
Irreversible • Antibiotics (penicillin and sulfa drugs)
• Inhibit enzymes essential to the life

Inhibitors processes of bacteria

• Interfere with transpeptidase in
penicillin
• Transpeptidase - Enzyme that is
important in bacterial cell wall
construction
Irreversible Inhibitor
- Cyanide Ion
• Extremely toxic and acts rapidly
• Interferes with the operation of an iron-
containing enzyme called cytochrome
oxidase
• Forms a stable complex and does not allow
the enzyme to function properly
• Results in cessation of cellular
respiration and causes death in a
matter of minutes
Irreversible Inhibitor
- Cyanide Ion
(continued)
• Antidote must be administered
quickly
• Sodium thiosulfate converts the
cyanide ion to a thiocyanate ion,
which does not bind to the iron of
cytochrome oxidase
Irreversible Inhibitor
- Heavy Metal
Toxicity
• Caused by the ability of heavy
metals, such as mercury and lead,
to render the protein part of
enzymes ineffective
• Metals act by combining with the —
SH groups found on many enzymes

• Cause nonspecific protein

denaturation
 Irreversible Inhibitor -
Heavy Metal Toxicity
(continued)

• Mercury and lead poisoning can cause permanent neurological damage

• Treated by administering chelating agents, which combine with
metal ions and hold them very tightly
• Example - Ethylenediaminetetraacetic acid (EDTA)
• Chelates all heavy metals except mercury
• Calcium salt of EDTA is administered intravenously
• Calcium ions are displaced by heavy-metal ions
that bind to the chelate more tightly
• Lead–EDTA complex is soluble in body fluids and is
excreted in the urine
FOUR WIDELY
USED
PENICILLINS
Reversible Inhibitors
• Reversibly bind to an enzyme
• Can be removed from the enzyme by shifting the established equilibrium

⎯⎯
→ EI
E + I ⎯
⎯
• Types
• Competitive inhibitors: Compete with substrate for binding at the active site
• Noncompetitive inhibitors: Bind to the enzyme at a location other than the
active site
 • Have molecular structures that are similar to the
normal substrate of the enzyme
• Example - Action of sulfa drugs on bacteria
• Folic acid is essential for the growth of

Competitive certain disease-causing bacteria

• Synthesized within the bacteria by a
process that requires p-aminobenzoic
Inhibitors acid
• Sulfanilamide resembles p-aminobenzoic
acid and competes with it for the active site
of the bacterial enzyme
Structural Relationships of Sulfanilamide, p-Aminobenzoic Acid, and
Folic Acid
THE BEHAVIOR OF COMPETITIVE INHIBITORS
Competitive
Inhibitors
(continued)
• Action can be reversed by:
• Increasing substrate concentration
• Letting LeChâtelier’s principle operate

• E - Enzyme
• S - Substrate
• I - Competitive inhibitor
Noncompetitive
Inhibitors
• Bear no resemblance to the enzyme substrate
• Interaction between the enzyme and the inhibitor
causes the 3-D shape of the enzyme and its active
site to change
• Substrate is improperly bound to the active
site
• Prevents catalytic groups of the active
site from participating in reaction
catalysis
• Substrate concentration does not affect inhibitor
action
Exercise
• Which of the following, when bound to an
enzyme, will increase the enzyme activity?
1. Competitive inhibitor
2. Noncompetitive inhibitor
3. Negative modulator
4. Positive modulator
Regulation of Enzyme Activity - Activation of Zymogens

• Zymogens (proenzymes): Inactive precursors of enzymes

• Some enzymes that would degrade the internal structures of the cell
are stored as inactive zymogens
• Released when required
• Activated at the location where the reaction occurs
• Involves the cleavage of one or more peptide bonds of the zymogen
• Example - Synthesis of trypsinogen
trypsinogen + H 2 O ⎯⎯⎯⎯⎯
→ trypsin + hexapeptide
enteropeptidase
EXAMPLES OF
ZYMOGENS
 Regulation of Enzyme Activity -
Allosteric Regulation

Refers to the
Feedback
control of an
inhibition
allosteric enzyme
Allosteric enzyme: Enzyme with a
quaternary structure whose
activity is changed by the binding Process in which the end product
of modulators of a sequence of enzyme-catalyzed
• Modulators: Bind to an enzyme’s active site reactions inhibits an earlier step in
and alter its catalytic activity the process
• May increase (activators) or decrease
(inhibitors) activity
Allosteric Regulation
- Example
• Synthesis of isoleucine is a five-step process
• Threonine deaminase (catalyzes in the first
step) is subject to inhibition from isoleucine
(final product)
• Isoleucine is a noncompetitive inhibitor,
which binds to an allosteric site and not an
active site
• Exerts an inhibiting effect on the
enzyme activity
• Reaction slows as its concentration
increases and no excess isoleucine is
produced
Regulation of
Enzyme Activity -
Genetic Control
• Synthesis of all proteins, including
enzymes, is under genetic control by
nucleic acids
• Example
• Enzyme induction: Synthesis of an
enzyme in response to a cellular need
• Example - Synthesis of -
galactosidase

• Allows organisms to adapt to

environmental changes
 SUMMARY
CHART OF
ENZYME
INHIBITORS
 • A regulator is a _____ inhibitor if it is involved in
_____ modulation
1. competitive; negative

Exercise 2. competitive; positive

3. noncompetitive; negative
4. noncompetitive; positive
 • Changes in blood serum concentrations of
specific enzymes can be used to:
• Detect cell damage and uncontrolled cell
growth

Enzymes in • Suggest the site of damage or cancer

• Estimate extent of cell damage

Clinical • Determined by the magnitude of

increase in serum concentration
above normal level

Diagnosis • Measurement of enzyme concentrations in

blood serum has become a major diagnostic
tool in diagnosing diseases of the heart, liver,
pancreas, prostate, and bones
DIAGNOSTICALLY USEFUL ASSAYS OF
BLOOD SERUM ENZYMES
Exercise
• Which of the following is NOT a form of
enzyme regulation?
1. Feedback control
2. Proenzymes
3. Allosterism
4. Hemotropism
Isoenzymes
• Slightly different forms of the same enzyme produced by different tissues
• All forms catalyze the same reaction but their molecular structures slightly differ and their locations may vary
• Example - Lactate dehydrogenase (LDH), whose serum levels are used in the diagnosis of:
• Anemias involving the rupture of red blood cells
• Acute liver diseases
• Congestive heart failure
• Muscular diseases, such as muscular dystrophy
Exercise
• Isoenzyme levels in the blood serum can
be used as a diagnostic tool because _____
1. more than one of these answers is
correct
2. these levels absolutely identify the
cause of a health issue
3. different tissues produce slightly
different forms of the same enzyme
4. isoenzymes are manufactured
in response to infection