*Enzyme applications#
Enzyme Inhibitors
Regulator or feedback - used to control a sequence of reactions -
reaction product may block initial enzyme.
Enzyme Inhibitors
Enzyme Inhibitors
Drugs as Enzyme Inhibitors
Drug mimics some natural chemical to block enzyme.
Drugs as Enzyme Inhibitors
Drug mimics some natural chemical to block enzyme.
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
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
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
