Roy method: Substrate: Thymophthalein Monophosphate CHEM. INHIBITION: 1. TARTRATE ( - prostatic ACP) 2. COPPER AND FORMALDEHYDE (- RBC ACP)

ENZYMOLOGY Flashcards by JA Unknown

Uses olive oil or triolein as substrate ( TAG + H2O in the presence of LIPASE will produce GLYCEROL and FATTY ACIDS) FA will be SUBSEQUENTLY TITRATED

Cherry-crandall method (LIPASE DETERMINATION)

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Measures the rate of clearing as fats are hydrolyzed by LPS

TURBIDIMETRIC METHOD ( LPS DETERMINATION)

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Based on coupled reactions with Glycerol Kinase and Peroxidase ( detection of glycerol)

Colorimetric methods (LPS determination)

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Reference value for LPS

<38 U/L

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Significance:

Hepatolbilliary disorders, Alcoholic Liver Disease, Microsomal induction by drugs and alcohol; also used to ID the source of increased ALP

GAMMA GLUTAMYL TRANSFERASE (GGT)

>GGT and 5NT = hepatolbilliary disorders Normal GGT and >5NT = Biliary obstruction

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Best enzymatic marker for ALCOHOLISM

GGT

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Clinical significance:
INTRAHEPATIC CHOLELSTASIS

also use to ID source of elevated ALP

5` Nucleotidase

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Major source of ACP

Prostate, RBCs

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Significance of ACP

Prostatic Cancer, hyperplasia, RAPE CASES

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ACP determination

Roy method:
Substrate: Thymophthalein Monophosphate
CHEM. INHIBITION:

TARTRATE ( - prostatic ACP)
COPPER AND FORMALDEHYDE (- RBC ACP)

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Aldolase significance

Skeletal muscle disorders

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Enzyme responsible for BP regulation

Angiotensin converting enzyme (ACE)

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Glycogen phosphorylase significance

AMI

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Pancreatic marker
Increased in acute pancreatic insufficiency
And decreased in chronic pancreatic insufficiency

Trypsin

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Miscellaneous enzymes that are only significant when increase

ACP
ALDOLASE
GUTAMATE DH
GLUTARHIONE-S-T
ACE
GLYOCGEN PHOSPHORYLASE
TRYPSIN

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Miscellaneous enzymes that are only significant when decrease

Chymotrypsin/Elastase 1
G-6-PD
PK
pseudo ChE

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Clinical significant if decreased in the following conditions

Chronic pancreatic insufficiency
Steatorrhea
Undigested muscle fibers

Chymotrypsin/Elastase 1

Miscellaneous enzyme decreased in drug induced hemolytic anemia

G-6-PD

Clinical significant of PYRUVATE KINASE

If decreased, in Hemolytic disorders such as Hemolytic anemia secondary to membrane abnormality. Congenital echinocytosis

Significance of Pseudo ChE

When decreased in SERUM, Pesticide poisoning, Liver disease, Genetic variance (prolonged response to muscle relaxants)

O T H L I L
1 2 3 4 5 6

Oxidoreductase
Trnasferases
Hydrolase
Lyases
Isomerase
Ligase

Biologic protein catalyst that accelerates biochemical reactions without being consumed or changed in composition

Enzyme

Catalytic mechanism

E + S <—-> ES complex <—-> P + E

• Substrate <Enzyme
• Enzyme> Substrate

Substrate- rate-limiting component

First-Order Kinetics

• Enzyme Enzyme

Zero-Order kinetics

2 ways to plot Enzyme Activity

Michaelis - menten Lineweaver-Burke plot

Measurements of enzyme activity or rate of velocity

IU - mmol/minute Katal - mol/second "Conversions: 1IU = 0.0167 Ukat = 16. 7 1 kat = 1.6 IU"

Factors that influence enzymatic reaction

pH Temperature Cofactors Inhinitors

Noncompetitive inhibitor

Binding site is *allosteric site* Addition of S *no effect* Low Vmax Same Km

Competitive inhibition

Binding site is *active site* Addition of S*reverses inhibition* Same Vmax Low Km

Uncompetitive inhibition

Binding site is *ES complex* Addition of S *increases the inhibition* Low Vmax and Km

Enzyme + Immunoglobulin

Macro enzyme (type 1)

Enzyme + no immunoglobulin substance

Macro enzyme (type 2)

Enzymes with macro forms

CK AMS LD AST ACP ALP LPS GGT

Creatinine kinase tissue sources

BRAIN HEART SKELETAL MUSCLES

Creatinine kinase pronounced elevation

**Duchenne's muscular dystrophy**, polymyositis, Dermapolymyositis, Myocardial infarction

CK isoenzymes

CK1 CK2 *MAXRO-CK* CK3 *MI*

CK determination tests

Tanzer-Gilvarg Oliver-Rosalki

LDH tissue sources

LD1- Heart, RBCs, Renal Cortex LD2 - same as LD1 LD3 - Lungs, Lymphocytes, Spleen, Pancreas LD4- liver, skeletal muscle LD5 - same as LD5

LD6

Alcohol dehydrogenase

CK mild to moderate elevation

Acute agitated psychosis, alcoholic myopathy, severe exercise, delirium tremens, severe ischemic Injury, pulmonary infarction, intramuscular injection, hypothyroidism