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Medical use of
Enzymes
Prepared by: Mohammad Reza Abdullahi
Master student in IIUM, Malaysia
Content
• Localization of enzymes
• Diagnosis and prognosis of disease
• Liver enzyme
• Pancreas enzyme
• Muscle enzyme
• Enzymes as analytic reagents
• Enzymes as therapeutic agents
Clinical
use of
enzymes
Diagnosis and prognosis of
disease
Analytic reagents
Therapeutic agents
Localization of enzymes
• intracellular
• Lysosome
• Nucleous
• Cytosol
• Mithochondria
• Extracellular
• Are secreted and function out from the cell
• Mainly digestive enzymes (alfa amylase)
Enzymes for diagnosis and
prognosis of disease
Liver
enzymes
• Aspartate aminotransferase
• Alanine aminotransferase
• Gamma glutamyl transferase
• Alkaline phosphatase
• 5’- nucleotidase
Aspartate aminotransferase
• AST/ SGOT/ GOT
• Requires pyridoxal 5’ phosphate
• Tissue sources
• Widely distributed
• Cardiac muscle, liver and skeletal muscle (higher concentration)
• Kidney, pancreas and erythrocyte (lower concentration)
Clinical significance
Hepatocellular
disorder
Viral hepatitis
Cirrhosis
Skeletal
muscle
disorder
Muscular dystrophy
Cardiac
disease
AMI
Congestive heart failure
Alanine aminotransferase
• ALT/ GPT/ SGPT
• Tissue sources
• All tissue
• Higher concentration in Liver
Clinical significance
Evaluation of
hepatic disorder
ALT is higher than
AST in acute
inflammation
condition
Longer half life
than AST
Îł-Glutamyltransferase
• GGT
• catalyze the transfer of amino acids from one peptide to another
amino acid or peptide
Tissue sources
• Kidney, brain, prostate, pancreas
and liver
Clinical significance
• Biliary tract obstruction
• Medications (warfarin,
phenobarbital and phenytoin)
• Alcohol consumption
• GGT activity is normal during
pregnancy and in patients with
bone disorders, conditions that
exhibit elevated ALP activity
Alkaline phosphatase
• ALP
• catalyze the hydrolysis of various phosphomonoesters at an alkaline
PH
• Require Mg++
• Tissue sources
• Bone, liver, intenstin, spleen,
• Placenta, kidney
Clinical
significance
• Biliary tract obstruction
• Bone disease
• Physiologically increases in pregnancy, infant
and children
5’-nucleotidase
• A phosphatase that acts only on nucleoside 5’-phasphates, such as
AMP and adenylic acid, releasing inorganic phosphate.
• Mainly increase in hepatobiliary disease
Pancreatic
enzymes
•Amylase
•Lipase
Amylase
• AMS
• Catalyze hydrolysis of α, 1-4 glycosidic bonds in starch and glycogen to
produce glucose, maltose and intermediate chains called dextrins which
contain α, 1-6 branching linkages.
• Require Calcium
• Normally seen in urine
• Tissue sources
• Pancreas
• Salivary gland
• Clinical significance
• Acute pancreatitis
• Mumps
Lipase
• catalyzes the hydrolysis of triglycerides to produce glycerol and fatty
acids
• Tissue source
• Pancreas
• Clinical significance
• Acute pancreatitis
• lipase activity increases 5 to 8 hours after the onset of symptoms, peak at 24
hours and a return to normal after 8 to 14 days.
Muscle
enzymes
• Creatine kinase
• Lactate dehydrogenase
• Glycogen phosphorylase
Creatine Kinase
• CK/ CPK
• catalyze the reversible phosphorylation of creatine to creatine
phosphate by ATP
• It is an enzyme found primarily in the heart and skeletal muscles
• Dimer with subunits B and M
Isoenzy
me
name
Compo
sition
Present in Elevated in
CK-1
Fast
moving
BB
Brain,prostate,GI
tract,lung,bladder,uteru
s,placenta
CNS diseases
CK-2
2% of
total
MB Myocardium/ Heart
Acute myocardial
infarction
CK-3
Slow
moving
MM
Skeletal muscle,
Myocardium
Clinical
significance
• AMI
• Muscular dystrophy
• Following MI, the CK-MB levels begin to rise
within 4 to 8 hours, peak at 12 to 24 hours and
return to normal levels within 48 to 72 hours
Lactate Dehydrogenase
• LD/ LDH
• catalyzes the interconversion of lactic to pyruvic acids. It is a
hydrogen-transfer enzyme that uses the coenzyme NAD+
• Tetramer (H and M)
Isoenzyme
name
Composition Electrophore
tic migration
Present in Elevated in
LDH 1
Heat
resistant
( H4) Fastest
moving
Myocardium,
RBC,kidney
myocardial
infarction
LDH2
Heat
resistant
(H3M1) Myocardium,
RBC,kidney
Kidney
disease,megalo
blastic anemia
LDH3 (H2M2) brain Leukemia,malig
nancy
LDH4
Heat labile
(H1M3) Lung,spleen Pulmonary
infarction
LDH5
Heat labile
Inhibited by
urea
(M4) Slowest
moving
Skeletal
muscle, Liver
Skeletal muscle
and liver
diseases
Clinical significance
The highest levels of total LDH are seen in pernicious anemia and
hemolytic disorders
Liver disorders, such as viral hepatitis and cirrhosis show slight
elevations of two to three times ULN
In AMI, LDH levels begin to rise within 12 to 24 hours, reach peak levels
within 48 to 72 hours, and may remain elevated for 10 days
Skeletal muscle disorders and some leukemias contribute to increased
LDH levels
Glycogen phosphorylase
• Glycogen phosphorylase uses inorganic HPO4
2+ to split glucose from
the polysaccharide chains of glycogen
• The glucose 1-phosphate so formed can be used for ATP synthesis in
muscle or converted to free glucose in the liver
• Requires pyridoxal phosphate as coenzyme
• Lack of glycogen phosphorylase result in Mc Ardle disease (glycogen
storage disease type 5) which causes muscle cramp and muscle
damage due to inadequate energy supply.
Enzymes as Analytic Reagents
• Measurement of substrates, drugs and enzyme activity
• Specific for substrate (advantage than chemical method)
• Direct measurement of the substrate in the complex mixture
• Methods for substrate measurements
• End point (the substrate is completely converted to product before it is
measured)
• Two-point Kinetic method (a change in substrate concentration produced
during a fixed-time interval is measured)
Enzymes as therapeutic agents
• Transfusion of fresh blood or its active components in bleeding
disorders
• Oral administration of digestive enzymes in digestive diseases
• Administration of fibrinolytic enzymes (e.g. streptokinase)
• Cancer treatment
Reference
• Tietz Fundamentals of Clinical Chemistry and Molecular Diagnostics
2015
• Clinical Chemistry Techniques Principles Correlations, Michael
L.Bishop and others, 2010
• Clinical chemistry, fundementals and laboratory techniques, Donna
L.Larson 2017
Thank you

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Medical use of enzymes

  • 1. Medical use of Enzymes Prepared by: Mohammad Reza Abdullahi Master student in IIUM, Malaysia
  • 2. Content • Localization of enzymes • Diagnosis and prognosis of disease • Liver enzyme • Pancreas enzyme • Muscle enzyme • Enzymes as analytic reagents • Enzymes as therapeutic agents
  • 3. Clinical use of enzymes Diagnosis and prognosis of disease Analytic reagents Therapeutic agents
  • 4. Localization of enzymes • intracellular • Lysosome • Nucleous • Cytosol • Mithochondria • Extracellular • Are secreted and function out from the cell • Mainly digestive enzymes (alfa amylase)
  • 5. Enzymes for diagnosis and prognosis of disease
  • 6. Liver enzymes • Aspartate aminotransferase • Alanine aminotransferase • Gamma glutamyl transferase • Alkaline phosphatase • 5’- nucleotidase
  • 7. Aspartate aminotransferase • AST/ SGOT/ GOT • Requires pyridoxal 5’ phosphate • Tissue sources • Widely distributed • Cardiac muscle, liver and skeletal muscle (higher concentration) • Kidney, pancreas and erythrocyte (lower concentration)
  • 9. Alanine aminotransferase • ALT/ GPT/ SGPT • Tissue sources • All tissue • Higher concentration in Liver
  • 10. Clinical significance Evaluation of hepatic disorder ALT is higher than AST in acute inflammation condition Longer half life than AST
  • 11. Îł-Glutamyltransferase • GGT • catalyze the transfer of amino acids from one peptide to another amino acid or peptide
  • 12. Tissue sources • Kidney, brain, prostate, pancreas and liver Clinical significance • Biliary tract obstruction • Medications (warfarin, phenobarbital and phenytoin) • Alcohol consumption • GGT activity is normal during pregnancy and in patients with bone disorders, conditions that exhibit elevated ALP activity
  • 13. Alkaline phosphatase • ALP • catalyze the hydrolysis of various phosphomonoesters at an alkaline PH • Require Mg++ • Tissue sources • Bone, liver, intenstin, spleen, • Placenta, kidney
  • 14. Clinical significance • Biliary tract obstruction • Bone disease • Physiologically increases in pregnancy, infant and children
  • 15. 5’-nucleotidase • A phosphatase that acts only on nucleoside 5’-phasphates, such as AMP and adenylic acid, releasing inorganic phosphate. • Mainly increase in hepatobiliary disease
  • 17. Amylase • AMS • Catalyze hydrolysis of α, 1-4 glycosidic bonds in starch and glycogen to produce glucose, maltose and intermediate chains called dextrins which contain α, 1-6 branching linkages. • Require Calcium • Normally seen in urine • Tissue sources • Pancreas • Salivary gland • Clinical significance • Acute pancreatitis • Mumps
  • 18. Lipase • catalyzes the hydrolysis of triglycerides to produce glycerol and fatty acids • Tissue source • Pancreas • Clinical significance • Acute pancreatitis • lipase activity increases 5 to 8 hours after the onset of symptoms, peak at 24 hours and a return to normal after 8 to 14 days.
  • 19. Muscle enzymes • Creatine kinase • Lactate dehydrogenase • Glycogen phosphorylase
  • 20. Creatine Kinase • CK/ CPK • catalyze the reversible phosphorylation of creatine to creatine phosphate by ATP • It is an enzyme found primarily in the heart and skeletal muscles • Dimer with subunits B and M
  • 21. Isoenzy me name Compo sition Present in Elevated in CK-1 Fast moving BB Brain,prostate,GI tract,lung,bladder,uteru s,placenta CNS diseases CK-2 2% of total MB Myocardium/ Heart Acute myocardial infarction CK-3 Slow moving MM Skeletal muscle, Myocardium
  • 22. Clinical significance • AMI • Muscular dystrophy • Following MI, the CK-MB levels begin to rise within 4 to 8 hours, peak at 12 to 24 hours and return to normal levels within 48 to 72 hours
  • 23. Lactate Dehydrogenase • LD/ LDH • catalyzes the interconversion of lactic to pyruvic acids. It is a hydrogen-transfer enzyme that uses the coenzyme NAD+ • Tetramer (H and M)
  • 24. Isoenzyme name Composition Electrophore tic migration Present in Elevated in LDH 1 Heat resistant ( H4) Fastest moving Myocardium, RBC,kidney myocardial infarction LDH2 Heat resistant (H3M1) Myocardium, RBC,kidney Kidney disease,megalo blastic anemia LDH3 (H2M2) brain Leukemia,malig nancy LDH4 Heat labile (H1M3) Lung,spleen Pulmonary infarction LDH5 Heat labile Inhibited by urea (M4) Slowest moving Skeletal muscle, Liver Skeletal muscle and liver diseases
  • 25. Clinical significance The highest levels of total LDH are seen in pernicious anemia and hemolytic disorders Liver disorders, such as viral hepatitis and cirrhosis show slight elevations of two to three times ULN In AMI, LDH levels begin to rise within 12 to 24 hours, reach peak levels within 48 to 72 hours, and may remain elevated for 10 days Skeletal muscle disorders and some leukemias contribute to increased LDH levels
  • 26.
  • 27. Glycogen phosphorylase • Glycogen phosphorylase uses inorganic HPO4 2+ to split glucose from the polysaccharide chains of glycogen • The glucose 1-phosphate so formed can be used for ATP synthesis in muscle or converted to free glucose in the liver • Requires pyridoxal phosphate as coenzyme • Lack of glycogen phosphorylase result in Mc Ardle disease (glycogen storage disease type 5) which causes muscle cramp and muscle damage due to inadequate energy supply.
  • 28. Enzymes as Analytic Reagents • Measurement of substrates, drugs and enzyme activity • Specific for substrate (advantage than chemical method) • Direct measurement of the substrate in the complex mixture • Methods for substrate measurements • End point (the substrate is completely converted to product before it is measured) • Two-point Kinetic method (a change in substrate concentration produced during a fixed-time interval is measured)
  • 29.
  • 30. Enzymes as therapeutic agents • Transfusion of fresh blood or its active components in bleeding disorders • Oral administration of digestive enzymes in digestive diseases • Administration of fibrinolytic enzymes (e.g. streptokinase) • Cancer treatment
  • 31.
  • 32. Reference • Tietz Fundamentals of Clinical Chemistry and Molecular Diagnostics 2015 • Clinical Chemistry Techniques Principles Correlations, Michael L.Bishop and others, 2010 • Clinical chemistry, fundementals and laboratory techniques, Donna L.Larson 2017

Editor's Notes

  1. With concentrations up to 5000 times higher than in other tissues