ENZYMES IN MEDICINE

• Diagnostic indicators – the activities of many enzymes are

routinely determined in plasma ( rarely in tissue biopsies) for
diagnostic purposes in diseases of the heart, liver, skeletal muscle,
pancreas and other tissues - enzyme diagnostics

• Therapeutic agents – several enzymes are used as drugs; new

approach - enzymotherapy

• Diagnostic tools – use as chemicals in clinical laboratory assays

 ENZYMES IN CLINICAL DIAGNOSIS
secretory - produced by tissues (namely liver), acting in plasma –

prothrombin, plasminogen, cerruloplasmin, choline

esterase; lipoprotein lipase
Enzymes

intracellular – function intracellulary, have no physiological use in

plasma
- membrane bound – ALP, GMT
- cytosolic – ALT, AST, LD, MDH
- mitochondrial – AST, GMDH
- lysosomal - ACP

- tissue specific – glucose-6-phosphatase – liver

amylase – pancrease
LD1 – heart
• Healthy individuals - levels of intracellular enzymes fairly constant, low –
the rate of enzyme release from damaged cells into plasma balanced by the rate
of removal of enzyme protein from plasma

Physiological enzyme levels  reference values of the enzyme activities

(determined in clinical laboratory – each lab

has its own reference values)

• Elevated enzyme activity in the plasma – reflect tissue damage accompanied

by increased release of intracellular enzyme
Skeletal muscle during exertion – physiologically elevated levels of muscle
enzymes in plasma

• Many diagnostically important enzymes = isoenzymes – pattern of

isoenzymes in plasma (determined electroforetically)
– a means of identifying the damaged tissue
ALTERATION OF ENZYME PLASMA LEVELS

Increased values – increased cell membrane permeability

anoxia, disturbances of energy metabolism 

cytosolic enzymes – ALT, LD, CK

- cell necrosis  membrane-bound enzymes – ALP, GMT

mitochondrial enzymes – AST, GMDH
- induction of the enzyme synthesis  drugs – ALP, GMT

Decreased values – inhibition of the activity  drugs

- inhibition of the synthesis  cell damage, drugs
 Examples of enzymes commonly assayed for diagnostic purposes

Enzyme Location Cause of elevated plasma level

Acid phosphatase - ACP Prostate Prostatic cancer

Alkaline phosphatase – ALP Bone, liver Rickets, hypoparathyroidism,

osteomalacia, obstructive
jaundice, cancer of bone/liver

Alanine aminotransferase – ALT Liver (muscle, Hepatitis, jaundice, circulatory

heart, kidney) faillure with liver congestion

Aspartate aminotransferase – AST Heart, muscle, Myocardial infarction, muscle

red cells, liver damage, anemia, hepatitis,

circulatory faillure with liver
congestion

Amylase - AM Pancres Acute pancreatitis, peptic ulcer

-Glutamyl transferase – GMT Liver, kidney, Hepatitis, alcoholic liver

pancreas damage, cholestasis
Examples of isoenzymes commonly assayed for diagnostic purposes

Enzyme Location Cause of elevated plasma level

Creatine kinase – CK
CK-MB Heart Myocardial infarction
CK-MM Skeletal muscle Muscular dystrophy

Lactate dehydrogenase – LD
LD1 > LD2 Heart, kidney, Myocardial infarction, kidney
blood cells disease, megaloblastic anemia,

leukemia
LD2, LD3 Leukemia
LD5 Liver, muscle Liver disease, muscle damage
 ENZYMES IN THERAPY
• Substitution of missing production of digestive enzymes – digestive
enzymes – pepsin trypsin…

• Removal of deposits of death tissue or fibrin (e.g. in lungs, eyes),

treatment of skin defects – proteinases, nucleases, collagenase

• Acceleration of fibrinolysis in lungs embolization (activation of plasmin

and plasminogen) – streptokinase, urokinase

 ENZYMOTHERAPY
Orally administered enzymes – treatment of a variety disorders
- digestive, gastrointestinal, pancreatic
- inflammatory diseases, edema
- immune and autoimmune diseases
(arthritis, multiple sclerosis)
- viral diseases (herpes, AIDS)
- cancer

Mixtures of enzymes of plant and/or animal origin - proteinases, amylase,

lipase - administered as acidoresistent tablets

• Pancreatin – trypsin, chymotrypsin, lipase, amylase

• Wobenzym – pancreatic and plant proteolytic enzymes – trypsin,
chymotrypsin, papain (Carica papaya), bromelain (ananas) =
combination of enzymes with different specificity, pH optimum,
stability, interaction with inhibitors and antiproteinases
 multiple action
• Mechanism of resorption (transport of large macromolecules across
the intestinal barrier) – paracellular transport, receptor mediated
endocytosis and transcytosis

• Mechanism of action – interaction with plasma antiproteinases –

1-antitrypsin, 2-macroglobulin complexes

direct proteolytic action, degradation of adhesive

? molecules, secretion of cytokins (tranforming growth factor
TGF-), modulation of receptor function

not fully clarified

ENZYMES - USE IN LABORATORY ASSAYS
Enzymes isolated from different sources - used for determination of various
substances in the blood, plasma/serum and urine  enzyme methods

much more specific than chemical methods, the presence of relative

substances with similar chemical properties does not hinder

Components of commercial kits or diagnostic strips

- determination of glucose - glucose oxidase, peroxidase
cholesterol - cholesterol esterase, cholesterol oxidase
peroxidase,
urea – urease, ……. in blood, plasma, serum
- proof of glucose (glucose oxidase), …….. in blood or urine (strips)

Markes in the immunochemical analysis

- ELISA (=enzyme-linked immunoadsorbent assay) – peroxidase, alkaline
phosphatase
NUCLEOTIDES
Structure, Function
 NUCLEOTIDE STRUCTURE

Nucleotides

nitrogenous base + pentose + phosphate group(s)

purine ribose 1-3
pyrimidine deoxyribose
other (nicotinamide)

Nucleosides
 THE
THE NITROGENOUS
NITROGENOUS BASES
BASES
Purine bases
NH2 O
| adenine
adenine || guanine
guanine
C N C
N C N
HN C
CH CH
HC C C C
N N H2N N
N
H H

Pyrimidine bases
NH2 O O
| || CH3 ||
C C C
N CH HN C
HN CH

C C CH
CH O C CH
O N N O
H H N
H
cytosine
cytosine thymine
thymine uracil
uracil
 NUCLEOSIDE

• A sugar - base combination.

Base
Base

HOCH2 O -N-glycosidic
-N-glycosidic
H H linkage
linkage
H H
OH H
Sugar
Sugar
In
In this
this case
case
deoxyribose
deoxyribose
 O thymine uracil
CH3 O
C
HN C C H
HN C
C CH
O N C CH
O N
HOCH2 O
HOCH2 O
H H
H H H H
H H
OH H
OH OH
deoxythymidine
NH2 uridine
|
C
N CH

C CH
O N cytosine
HOCH2 O
H H
H H
OH OH
cytidine
 NH2 adenine guanine
| O
C N
N C C N
CH HN C
CH
HC C C C
N N N
H2N N
HOCH2 O
HOCH2 O
H H
H H H H
H H
OH H
OH OH
deoxyadenosine guanosine
OH
|
C N
N C hypoxanthine
CH
HC C
N N

HOCH2 O !
H H
H H
OH OH inosine
 NUCLEOTIDES
NUCLEOTIDES

5’-OH on the sugar of a nucleoside is converted into a

phosphate ester.

deoxyadenosine NH2
deoxyadenosinemonophosphate
monophosphate
|
(dAMP)
(dAMP) C
N
N C
CH
Each is named based on HC C
N
sugar and base name O N
and then the number of ||
-
O-P-O-CH
O
phosphates is indicated. | 2

H H
O- - H H
OH H
 ATP - adenosine triphosphate

adenine
phosphate chain NH2

O O O N
N
-
O P O P O P O

O- O- O- CH2 N N
O

OH OH

ribose
AMP
ADP
ATP
 NUCLEOTIDE FUNCTION

• Precursors of DNA, RNA - NTPs

• Energy transport - ATP
• Allosteric effectors of enzymes – ATP, ADP, AMP
• Covalent modification of enzymes – ATP
• Intracellular mediators (= second messengers) – cAMP, cGMP
• Coenzymes – NAD+, NADP+, FAD, CoA-SH
• Activated precursors of polysaccharaides, glycoproteins,
proteoglycans, phospholipids, glycolipids – UDPG, UDPGA,
UDPGal…, CDP-choline, CDP-diacylglycerol…
• Active groups (group transport) – SAM, PAPS
 NAD+
reactive O
site C
O-
NH2 nicotinamide
O P O N+
CH 2
O

O NH2
OH OH
N N
adenine
O P O
CH 2 N N
O - O

ribose
OH OH
 FAD
O
H3 C N
reactive site
NH

H3C N N O
H C H

H C OH
riboflavin
H C OH

H C OH
NH 2
H C H

O N N adenine
O P O CH 2 N N
O - O

ribose OH OH
 Coenzyme A

pantothenate phosphorylated
unit ADP
NH2
O O H CH3 O O N N
C-CH2-CH2-N-C-C-C-CH2 O P O P O
H HO CH3 O- O- CH 2 N N
H-N O
CH2-CH2

S
Sulfhydryl
O OH
H group
O P O-
O-
 CH2 OH CH2 OH

H O H H O OH
H H
OH H OH H
O H
OH
H OH H OH CH2 OH
H O
-D-glucose -D-glucose H
H
OH H
OH OH
H OH
 H O
H C O C
O
H C O C
O
2 O P -O- CH2
-
O
Non-polar tail
cAMP – cyclic adenosine monophosphate

cAMP
 NH2
C| N
N C
CH
HC C
N N

CH2 O
O H H
-
H H

O P O OH

O-

cAMP – cyclic adenosine monophosphate

-intracellular mediator, second messenger of hormonal signal tranduction
via adenylate cyclase cascade
- mechanism of action: allosteric effector
 O

O CH2-O-C
O O
C-O-CH
SCoA SCoA

CH2-O - P ~ P
HO HO HO

HO HO HO