branch of medicine which

deals with the study of

enzymes and their
importance in the
diagnosis and treatment
of diseases
CELLULAR
CATALYSTS
Biologic catalysts that
cause reactions in the
body to take place
1. Complicated type of protein in
terms of both structure and
function
2. Easily denatured with varying
molecular weight & mass
3. Amphoteric
4. Operate at high rates
5. Reactions catalyzed are frequently
REVERSIBLE
6. Usually operates with the assistance of
a non-protein COFACTOR
7. Synthesized in an inactive state
(ZYMOGEN/PROENZYME)
8. Changes in enzyme concentration in
tissue cells reflect changes in states of
health & of the tissue
1) COENZYME – non-protein organic biochemical
that takes part in the enzyme reaction
- Essential to the catalytic activity as a CO-
SUBSTRATE
- E.g. NAD, Pyridoxal phosphate
2) ACTIVATOR
- Inorganic ionic cofactor
- Usually metal ions (esp. divalent cations)
- Metabolic regulator of enzyme reaction
- E.g. Mg++, Na+, K+, Zn++
3) HOLOENZYME
- the combined enzyme & coenzyme
4) APOENZYME
- Enzyme without a cofactor
5) PROSTHETIC GROUP
- A coenzyme that cannot be removed from
its attachment to an enzyme using dialysis
- E.g. Pyridoxal phosphate in transaminase
reaction
6) METALLOENZYME
- Inorganic activators existing as a part
of the enzyme molecule
7) SUBSTRATE
- Substance acted upon by an enzyme & is
converted into a new substance
8) PRODUCT
- Substance derived from a transformed
substrate
FOUR EXISTING PROTEIN
STRUCTURES
A. PRIMARY STRUCTURE
- Refers to the sequence of
amino acids joined by peptide
bonds to form a polypeptide
chain
B. SECONDARY STRUCTURE
- Conformation of the segments
of polypeptide chain
- Alpha-helix ; beta-pleated
sheet
- Maintained by hydrogen bond
C. TERTIARY STRUCTURE
- Arises from the interactions
among side chains/groups of the
polypeptide chain
- Bended and folded structure
- Maintained by covalent disulfide
bond
The 2° and 3° structures: most
important configurations of an
enzyme
responsible for the conformation of
the active site
ACTIVE SITE – area on the
enzyme where the substrate
attaches & undergoes
transformation
D. QUARTERNARY STRUCTURE
- Separate bended & folded
structures are put together to form
a functional unit
- Examples: Hemoglobin – consists of
4 polypeptide units
Enzyme variants – LDH, Creatine
kinase
Based on catalytic activity
of an enzyme
The International Union of
Biochemistry (IUB) Enzyme
Commission categorized all
enzymes into six (6) classes
I. OXIDOREDUCTASES
 Catalyze oxidation-reduction reactions
 Older names: Dehydrogenases &
Oxidases

 Examples: Glucose oxidase

Cytochrome oxidase
Lactate dehydrogenase
Isocitrate dehydrogenase
II. TRANSFERASES
- Move an intact group of atoms from one
molecule to another (amine or phosphate
group)

- Examples: Aspartate aminotransferase

Alanine aminotransferase

Creatine kinase
 ATP + Creatine ADP +
Creatine PO4
III. HYDROLASES
 Splits molecules with water as part of
the reaction process
 Three (3) Groups:
A. Esterases: ACP, ALP, Lipase
B. Peptidases: Leucine aminopeptidase,
Pepsin
C. Glycosidases: Amylase, Amylo-1,6-
glucosidase
IV. LYASES
 Responsible for splitting molecules or
breaking of bonds (C to C; C to O; C to
N, etc.)
 Assayed in disorders of skeletal
muscles
 Examples: Aldolase
Glutamate
decarboxylase
Pyruvate decarboxylase
V. ISOMERASES
 Responsible for the conversion of one
isomer to another
 Transformation from:
Cis to Trans
L to D forms
Aldehyde to Ketone
 All reactions are reversible
 Example: Glucose PO4 Isomerase
VI. LIGASES
Enzymes causing bond
formation between two
molecules to form a larger
molecule
Examples: Ligase
Amino Acyl t-RNA
synthetase
1. Type of reaction catalyzed
Example: Oxidation of Glucose = Glucose
oxidase
2. Suffix “ASE” added to the name of
substrate
Examples: Urea = Urease
Uric acid = Uricase
Phosphate esters = Phosphatase
Lipid = Lipase
3. EMPIRICAL Name
Examples: Trypsin, Pepsin
4. STANDARD SYSTEM
- Formulated by IUB and IUPAC
A. Systematic Name
 Describes the nature of the reaction
catalyzed
 Numerical code designation prefixed
w/ the letters E.C.
 Examples: E.C. 3.1.3.1 = ALP
E.C. 3.1.3.2 = ACP
1st digit = denotes the class of the
enzyme
2nd digit = sub-class of the enzyme
3rd digit = subsub-class
4th digit = specific serial number
 This approach removes all ambiguity
about the enzyme’s identity
B. TRIVIAL NAME
 a.k.a Non- specific, Practical name,
Working name
 A simplification of the systematic name
 Uses acronyms and abbreviations
 Examples:
SGOT, SGPT
Several distinct forms of enzymes
Important in the diagnosis of
specificity
A. ISOENZYMES
- Multichained enzymes of similar
activity
- Appear in specific tissue, organ &
cell organelle of similar organisms
Example: Lactate Dehydrogenase (LDH)
 Contains H & M sub-units (polypeptide
chains)
H4 (LD1) – heart, RBC & renal tubules
H3M (LD2) – heart, RBC & renal tubules
H2M2 (LD3) – Lungs, Lymphocytes, spleen,
pancreas
HM3 (LD4) – Liver, skeletal muscles
M4 (LD5) – Liver, skeletal muscles
1. Charged molecules – as shown in
electrophoresis
2. Mobility in Ion Exchange Resin
3. Response to activation & inactivation
process
E.g. ACP (RBC) – not inactivated by 2%
Formalin
ACP (Prostate) – inactivated by 2%
Formalin
4. Relative substrate specificities
E.g. ACP (RBC) is less sensitive to
α-naphthyl PO4
5. Response to Inhibitors
E.g. ACP is inhibited by flouride,
heparin, oxalates
B. HETEROENZYME
- Enzymes of similar catalytic activity but
are specie specific
E.g. LDH of rabbits & humans
C. ALLOENZYME
 Genetically-transmitted enzyme
 Important in defining the biochemical
characteristics of an individual
 Forensic medicine & genetics
CLASSIFICATION OF ENZYMES IN
BLOOD
A. PLASMA-SPECIFIC ENZYMES
1. Generally secreted by the liver
2. Enzymes that exert their function in
plasma
3. Examples: Coagulation factors
Fibrinolytic factors
Complement system
B. NON-PLASMA SPECIFIC ENZYMES
1. No specific functions in plasma (lack of
activators or coenzymes)
2. Two Classes:
B.1 Enzymes of Secretion
- secreted in plasma at a high rate but
rapidly disposed off to excretory
channels
- low concentration in plasma is
maintained
 B.2 Enzymes Associated w/ Cellular
Metabolism
- carry out their functions within the
cells in which they are formed
- e.g. Creatine kinase = Heart
ACP = Prostate gland
I. UNILOCULAR ENZYME
- Found only in one location,
particularly the cell sap

BILOCULAR ENZYME
II.
- Found in the mitochondria & cell
sap
E + S ES P + E
Where: E = unchanged enzyme
(catalyst)
S = substrate
P = transformed substrate
ES = postulated enzyme-
substrate complex
Energy – activation energy
Molecular Compatibility –
commonness between Enzyme &
Substrate
Space Availability – # of
enzymes or substrates that can
be reacted
Specificity – refers to the
enzyme acting on a specific
Lock & Key – refers to the
active site being
complementary in shape & size
to the substrate
Induced Fit Model – the
enzyme changes in shape during
binding to accommodate the
substrate
1. TIME – rate of enzyme action
 If the catalytic activity of an
enzyme on the substrate is
fast = shorter reaction time
 The enzyme is freed & can act
again on the remaining
substrate
2. SUBSTRATE CONCENTRATION
– direct relationship
MICHAELIS-MENTEN CURVE –
shows the relationship of the
reaction velocity to the substrate
concentration
Two Phases:
a. First Order Kinetics
- Enzyme conc. is fixed; Substrate
conc. is varied
- Rate of reaction is almost directly
proportional to substrate conc. at
low values
- Rate of reaction is dependent on
substrate conc.
 Vmax
Reaction
velocity

Substrate concentration

 Faster reaction but it reaches a saturation point when all the enzyme
molecules are occupied.
 If you alter the concentration of the enzyme then Vmax will change too.

© 2007 Paul Billiet ODWS

46
b. Zero Order Kinetics
- Reaction rate is unaffected by
increased substrate concentration
- Dependent on enzyme concentration
- When maximum velocity is reached,
the rate of increase in velocity is
“O” (Zero order reaction)
3. Enzyme Concentration
Direct relationship
An increase in enzyme
concentration results to an increase
in the catalytic activity
4. Temperature
Optimum temperature – the
temperature w/c is considered
favorable for enzyme activity (30-
37°C or 37 – 40°C)
Q10 value – reaction rate is doubled
for every 10°C increase
50 – 60°C : enzyme undergoes
inactivation and denaturation
5. pH – Hydrogen Ion Concentration
Optimum pH – the point at w/c the
reaction rate is greatest
At pH 7.0 – 8.0, many enzymes show
maximum activity
Pepsin is active at 1.5 and ALP at
10.5
 Optimum pH values

Enzyme
activity
Trypsin

Pepsin

1 3 5 7 9 11

© 2007 Paul Billiet ODWS

pH
51
6. Activators
Bind the substrate to the active site
by forming ionic bridges
Orients the substrate so it is
attached to the enzyme in the correct
configuration
Examples
- Fe++ - Cl- - K+
- Ca++ - Br- - Na+
- Zn++ - NO3-
7. Inhibitors
- Decrease the rate of enzyme
reaction
- Functions:
Binds to the active site, blocks
access of the S to the E
(Competitive Inhibition)
Binds elsewhere on the E
causing change in shape that
interferes w/ S binding (Non-
Competitive Inhibition)
Binds w/ the E-S complex; no
product formed (Uncompetitive
Inhibition)
 TYPES of INHIBITION
a. Reversible Inhibition
- Inhibitors are possible removed from
the system; enzyme is fully restored
- Physical processes that remove
inhibitors: dialysis, gel filtration
b. Irreversible Inhibition
- Inhibitors covalently combine w/ the
enzyme
- Physical methods are ineffective in
separating inhibitors from the enzymes
1. Excess substrate – causes competition
between substrate molecules for a
single binding site
2. Product of reaction – may be an
inhibitor of the forward reaction
3. E-S complex does not break to yield
products
4. Chemical substances
E.g. Flouride, Oxalate, Heparin, Cupric
ion, Tartrate
8. Coenzyme concentration
 Required for the reaction to proceed
(Vitamins, Coenzyme A or B, NAD)
 Must be present in proper concentration
9. Prosthetic Group
- Coenzyme that firmly binds with the
enzyme
Disruption of the 3-dimensional
structure of the enzyme
molecule
Leads to the loss of enzyme
activity
May be reversed if the
reaction process has not gone
too far & if the denaturing
agent is removed