s

REVIEW ON ENZYME TERMINOLOGIES AND CONCEPTS

[All actions in the body are mediated by enzymes. Enzyme substrate with specific shape can bind to an active site]
are protein catalase – increased, they speed-up the rate of
reactions without being change in overall process]

[Enzyme are task to do specific actions come in contact with

Substrate and bind together. Substrate will fit to the enzyme
and transforms and producing new products]

[Enzymes substrate complex - substrate come contact with

enzymes (1) Substate is specific for certain enzymes. Active
site – cavity, where the reaction takes place; substrate
interacts w/enzyme. Allosteric site – site other than the
active site; another regulatory wander will bind to the enzyme
it can be regulatory molecule, inhibitor. When the substate
come in contact with active it will now form enzyme/substrate
com plex – enzyme slightly change its shape as the
substrate binds & forming products. Product are form, leave
the active site will go to the specific task. Substate –
compound which the enzyme works & speed up the reaction,
acted upon enzyme. Apoenzyme – protein portion of the
enzyme (inactive) come in contact w/ Cofactor – non-
proteins parts of the enzyme, help in reaction (activator).
Protein is still inactive needs a cofactor to be an Active
enzyme. 2 types of Cofactors (1) Metallic /Non-metallic (2)
Organic /Inorganic. Activators – type of cofactor that are
inorganic in nature & derived from dietary minerals.
Coenzyme – organic cofactor. Ex. of activator – Chloride,
magnesium. Ex of Coenzyme – NAD, NADH. When an
apoenzyme come in contact with cofactor it will now form
Holoenzyme – active. Apoenzyme cannot be activated w/o
the presence of the cofactor, and cofactor cannot perform
the enzyme reaction w/o the presence of apoenzyme. Active
enzyme – capable of binding to a substrate and eliciting a
rection. Isoenzyme – the same function it exist in diff. form
either found in diff tse sources or can have diff. properties
such as diff. electrophoretic mobility, solubility, resistance,
they have the same function of CK]
 LOCK AND KEY MODEL

[Not all substrate can fit in certain enzyme also enzyme.

Enzyme have predetermined shape for the active site, only a

MONTEMOR, DJ 1
 INDUCED FIT MODEL

[Allow minor changes to accommodate the

substate]
NOMENCLATURE
[enzyme is given name derived from the action they
catalyze or type of compound they act]
 Systematic Name – Substate they act on &
reaction catalyze or including the cofactor
 Recommended Name – shorter version of
systematic name, ex Creatine Kinase (CK)

[One enzyme have systematic name, recommended name

& EC number]
Enzyme Commission (EC)

[ O – T – H - L – I – L]

 Suffix -Ase = Enzyme [NOT ALL ENZYME END WITH

(ASE) SOME IN DIGESTIVE ENZYME END WITH
(IN)]
 Ex, Glucose oxidase - Oxidation reaction, Substrate
= Glucose
1. OXIDOREDUCTASES - Oxidation-reductions [link
to one another requires Coenzyme either: oxidize or
reduce as the substrate reaction]

2. TRANSFERASES - Transfer of functional group

[2 Types (1) Transaminase – Transfer of an Amino
group of

MONTEMOR, DJ 2
 substrates (2) Kinase - Subtype of Transferase;
Catalyzes Phosphate Group]

[!!ENZYMES ARE PROTEIN, BUT NOT ALL PROTEIN ARE

ENZYME]
3. HYDROLASES - Removal of water to break H  Temperature
bonds [enzyme catalyze hydrolysis reaction, addition of o Protein dentures at 40OC, also enzyme
water molecule to the bond to cause it to break; central o Destroy at 60OC
to the process of digestion] o High temp high kinetic energy causes increased in
activity. higher the temp the higher the activity but
there is optimum temperature (temp at its
maximum- exceeds cause damage)
 pH
o Depends on the enzyme (Most enzymes optimum
activity at 7-7.5 pH, there are also some enzymes
require specific enzyme)
 Substrate Concentration
o Enzyme not consumed in the reaction they catalyzed
o The enzyme activity increases w/ increase enzyme
concentration
o The more substrate the more enzymatic
 Enzyme Concentration
 FIRST-ORDER KINETICS
o Reaction rate is directly proportional to
substrate concentration [maximum reaction
4. LYASES - Addition of a group to a double bond or rate, depends on substrate concentration. If it
removal of a group to form a double bond [does reaches maximum level]
not involve hydrolysis/oxidation reaction]  ZERO-ORDER KINETICS
o Reaction rate depends only on enzyme
concentration [the more enzyme the more
reaction will occur]
INHIBITORS
[Inhibitors - interfere; not typically bad needed for regulation
(ex. need a enzymatic reaction to occur may kailangan na
process in the body, if it reaches its goal. If too much can
cause damage)]

5. ISOMERASES - Rearrangement of atoms [mirror

reaction]

6. LIGASES - Reactions involving bond formation

coupled with ATP hydrolysis [energy hydrolysis]
 Competitive inhibition

[Competitive Inhibitor – binds to the activate site and

compete w/ the substrate for active site. Reversible type – if
there is more substrate than inhibitors .If inhibitor (blocking

MONTEMOR, DJ 3
the active site) come contact w/active site it will interfere
leading to NO REACTION]

MONTEMOR, DJ 4
 o A competitive inhibitor is another woman
(home wrecker!). She is competing with the
substrate for the enzyme's active site. If the
competitive inhibitor binds to the enzyme,
she will block the active site and prevent a
chemical reaction from occurring with the
substrate. Hence no product will be formed.
 Non- Competitive Inhibition

[Inhibitor will not compete w/the substrate with active site,

but it will look for another site called Allosteric site. Bind to
allosteric site in which cause CHANGES. Irreversible type –
if the inhibitor will already destroy part of enzyme]

o A non-competitive inhibitor is another man.

He is not competing with the substrate for
the active site. Instead, he will bind to an
alternate allosteric site (.e. the rear end) if
the non-competitive inhibitor binds to the
enzyme, it causes a conformational change in
the shape of the active site, such that the
active site loses its affinity for the substrate
(i.e. the active site becomes flaccid around
the substrate). Hence no product will be
formed.
 Uncompetitive Inhibitor
o Binds to enzyme substrate complex
o Even increased amount of substrate it will not be
affected. The more substrate bind to enzyme the
more it bind to enzyme substrate complex

MONTEMOR, DJ 5
 ENZYMES OF CLINICAL SIGNIFICANCE
CK-MM + lipoproteins
GENERAL CONCEPT!  Isoenzymes:
 Enzymes are found in certain tissue sources (ex. o CK-BB, CK-MB, CK-MM (major isoenzyme) [CK- MM
enzyme found in the liver - whatever the tissue sources – greatest concentration in normal individual]
clinical significance. ALP found in the liver if damage in o 2 Subunits: B & M
the liver this enzyme leaks out bloodstream. o CK-Mi: bound to exterior surface of the
inner mitochondrial membranes (minor)
CREATINE KINASE (CK) o Macro CK: CK-BB + IgG
 CREATINE KINASE - Recommended name
 E.C.2.7.3.2
 ATP: Creatine N- phosphotransferase- Systematic
name
 Reaction Catalyzed:

[Every time muscles move it requires energy. Transfer of 1

phosphate molecule = diphosphate]
 Tissue sources:
o Skeletal muscles, heart muscle, brain tissues
[significant tissue source] [the significant tissue
source of enzyme will be the diagnostic significance
of the enzyme]
o Bladder, placenta, GIT, thyroid, uterus,
kidney, lung, prostate, spleen, liver, and
pancreas
 Function:
o Associated with ATP regeneration in
contractile or transport system
o Predominant in muscle cells
ENZYME TISSUE CONDITION
Heart Myocardial infarction
Skeletal Skeletal muscle disorder
muscles
CK-MM Muscular dystrophy
Polymyositis
Hypothyroidism
Malignant hyperthermia
Physical activity
Intramuscular injection
Heart Myocardial infarction
Skeletal Myocardial injury
muscle
Ischemia
Angina
Inflammatory heart
CK-MB disease
Cardiac surgery
Duchenne-type muscular
dystrophy [highest
elevation of CK]
Polymyositis
Malignant hyperthermia
Reye's syndrome
Rocky Mountain spotted
fever
Carbon monoxide
poisoning
Brain Central nervous system
shock
Bladder Anoxic encephalopathy
CK-BB Lung Cerebrovascular accident
Prostrat Seizure
e
Uterus Placental of uterine
Colon Carcinoma
Stomac Reye's syndrome
h
Thyroid Carbon monoxide
poisoning
Malignant hyperthermia
Acute and chronic renal
failure
MONTEMOR, DJ 6
GENERAL CONCEPT!!
 Physiologic increased - normal occurrence in the
body not disease related ex. exercise before blood
collection
 Pathologic - disease state (ex. AMI, old patients)
 Most specific & sensitive enzyme marker – CK
 Gold standard in AMI – Troponin (protein not enzyme)
MYOCARDIAL INFRACTION
RISE 4-8 hours
PEAK 12-24
hours
NORMALIZE 48-72
hours
 Protein Electrophoresis

[Isoenzyme have the same function but can be found in

different tissue sources with different properties] [the
numbering of isoenzyme is based from their
electrophoretic pattern; #1 most anodal/migrates
furthest]
 Assay
o Forward reaction: TANZER-GILVARG METHOD
 Coupled enzyme: PK-LD-NAD+
 Optimal pH: 9.0
 Reaction catalyzed:

o Backward reaction: OLIVER-ROSALKI METHOD

[commonly used coz 2x faster than forward]
 Coupled enzyme: Hexokinase -G6PD-NADP
 Optimal pH: 6.8
 Reaction catalyzed:

 Sources Of Error:
o Hemolysis [Adenylate Kinase (AK) – enzyme
release from RBC to hemolyze sample, falsely
elevate CK levels]
o Muscle activity prior to collection
LACTATE DEHYDROGENASE (LD/LDH)
 LACTATE DEHDROGENASE
 E.C.1.1.1.27
 L-Lactate: NAD+ Oxidoreductase – Systematic Name
 REACTION CATALYZED:

 Coenzyme: NAD+
 Principle:
o Catalyzes the interconversion of lactic
and pyruvic acids
 Tissue Sources:
o Widely distributed in the body heart, liver,
skeletal muscle, kidney, RBC, lung,
smooth

MONTEMOR, DJ 7
 muscle, brain [least specific coz widely distributes  Substrate: Pyruvate
in the body]  Endpoint: Lactate
 Isoenzyme  Optimal pH: 7.1-7.4
o 2 Subunits: H & M
 LD1: (HHHH) H4(anodal)
 LD2: (HHHM) H3M1
 LD3: (HHMM) H2M2
 LD4: (HMMM) H2M3
 LD5: (MMMM) M4
 LD6: Alcohol dehydrogenase (minor) [seen
in px. w/ Atherosclerotic cardiovascular
disease]
 LDX: (minor) Human
 LD FLIP PATTERN
o Normal: LD 2>1>3>4>5
o AMI: LD 1>2>3>4>5 [also seen px.
severe hemolysis]
ISOENZYME TISSUE DISORDER
LD-1(HHHH) Heart Myocardial infarct
and Red blood Hemolytic anemia
LD-2 (HHHM) cells
Renal cortex Megaloblastic
anemia
Acute renal infarct
Hemolyzed
specimen
LD-3 Lun Pulmonary
(HHMM) g embolism
Lymphocyte Extensive
s pulmonary
Spleen pneumonia
Pancreas Lymphocytosis
Acute pancreatitis
Carcinoma
LD-4 Liver Hepatic injury or
(HHMM) inflammation
and Skeletal Skeletal muscle
LD-5 muscle injury
(MMMM)

MYOCARDIAL INFRACTION
RISE 12-24
hours
PEAK 48-72
hours
NORMALIZE 10 days
 ELCTROPHOREIS

 Isoenzyme Assay
o Fluorimetry or Colorimetry
 Substrate: alpha-hydroxybutyrate

 Assay
o Backward reaction: WROBLEUSKI LA DUE
METHOD

MONTEMOR, DJ 8
 o Forward reaction: WACKER METHOD
 Substrate: Lactate
 Endpoint: Pyruvate
 Optimal pH: 8.3-8.9
 Sources of Error
o Hemolysis
o LD5: most heat labile
ASPARTATE AMINOTRANSFERASE (AST)
 ASPARTATE AMINOTRANSFERASE
 E.C.2.6.1.1
 L-Aspartate: 2 oxoglutarate aminotransferase
 OLD NAME: SGOT/GOT
o Serum glutamic oxaloacetic transaminase
 Reaction Catalyzed:

 Coenzyme:
o Pyridoxal phosphate/Vitamin B6
 Principle:
o Transfer of an amino group between
aspartate and a ketoacids to form
oxaloacetate and glutamate
 Substrate:
o Aspartate
 Tissue Sources:
o Heart and liver
o skeletal muscles, kidney, pancreas, RBC
 Clinical Significance:
o Heart and liver
o skeletal muscles, kidney, pancreas, RBC
o Acute Myocardial Infarction
o Hepatocellular disorders [Liver disease
can be divided into (1) Hepatocellular (2)
Hepatobiliary disorder. AST significantly
increased in Hepato compared to Hepatobila.
AST is increased in all liver dse]
 viral hepatitis, liver cirrhosis
o Muscular dystrophies
o Others
 pulmonary diseases
 inflammatory conditions
MYOCARDIAL INFRACTION
RISE 6-8 hours
PEAK 24 hours
NORMALIZE 5 days
 Assay:
o KARMEN METHOD
o Principle: catalyzes the reduction of
oxaloacetate to malate with the
oxidation of NADH
o Indicator: malate dehydrogenase (MD)
o Optimal pH: 7.3-7.8
o Abs: 340 nm

 Source of Error:
o Hemolysis

MONTEMOR, DJ 9
 ALANINE AMINOTRANSFERASE (ALT) o Liver, Bone, Placenta, Intestine
[Liver specific enzyme of transferases] o Spleen, Kidneys
 E.C.2.6.1.2
 L-Alanine: 2 oxoglutarate aminotransferase
 OLD NAME: SGPT/GPT
o Serum glutamic-pyruvic transaminase
 Reaction Catalyzed:

 Coenzyme:
o Pyridoxal phosphate/Vitamin B6
 Principle
o Transfer of an amino group between alanine
and a ketoacids to form pyruvate and
glutamate
 Substrate:
o Alanine
 Tissue Sources:
o Liver
o Heart, skeletal muscles, kidney, pancreas, RBC
 Clinical Significance:
o Hepatocellular disorders
 viral hepatitis, liver cirrhosis
o Hepatocellular > Hepatobiliary
 ALT > AST
 Source of Error:
o Relatively unaffected by hemolysis
 Assay:
o Principle: catalyzes the reduction of
pyruvate to lactate with the oxidation of
NADH
o Indicator: lactate dehydrogenase (LD)
o Optimal pH: 7.3-7.8
o Abs: 340 nm

 DE RITIS RATIO [ratio between AST&ALT levels]

o <1 =Normal/Nonalcoholic
o >1/>2 = Alcoholic Liver Disease
ALKALINE PHOSPHATASE (ALP)
 E.C.3.1.3.1
 Orthophosphoric Monoester Phosphohydrolase
(Alkaline Optimum)
 Reaction Catalyzed: [hydrolases]

 Principle:
o Catalyze the hydrolysis of
various phosphomonoesters at
an alkaline Ph
 Activators: [type of coenzyme]
o Magnesium
o Zinc
 Tissue Sources

MONTEMOR, DJ 1
0
 Isoenzymes: o pregnancy, growing bones, old age healing
o Liver ALP, Bone ALP, Placental ALP, Intestinal bone fractures
ALP
o [LIVER, found in sinusoidal and bile canalicular
membrane; BONE - involved in
osteoblasts(bone formation)]
 Methods of Separation
o Electrophoresis
 Liver(anodal)-Bone-
Placental
Intestinal(cathodal)
o Heat Stability [before & after heating the serum
at 56OC for 10mins.]
 Placental (heat stable)-Intestinal-
Liver-Bone (<20%; heat labile) [PILB]
o Chemical Inhibition
 PHENYLALANINE inhibits
- Intestinal-Placental
 LEVAMISOLE inhibits
- Bone-Liver
 Abnormal Isoenzymes:[associated with neoplasm]
o Regan
 most heat stable
 migrates as Bone ALP
 inhibited by phenylalanine
o Nagao
 carcinoplacental ALP
 similarities with Placental ALP
 inhibited by phenylalanine & Leucine
 Clinical Significance:

[AST & ALT (both  in liver dse) – sensitive to

Hepatocellular disorder (ex. viral hepa) > Hepatobiliary;
ALP – slight increased in viral hepa, mark increased in
Hepatobiliary]
o Liver Diseases
 Hepatocellular < Hepatobiliary
o Bone Disorder
 Paget's Disease (osteitis deformans)
o Intestinal Disorders
o Placental Disorders
 Assay:
o Bowers and McComb
o Principle: calculation of ALP activity in
the molar absorptivity of p-nitrophenol
o Abs: 405 nm

o p-nitrophenyl-phosphate is
hydrolyzed to p-
nitrophenol
 Source of Error:
o Affected by hemolysis
o Should be run ASAP
 Physiologic causes of varying levels:

MONTEMOR, DJ 1
1
 ACID PHOSPHATASE (ACP)
 E.C.3.1.3.2
 Orthophosphoric Monoester Phosphohydrolase
(Acid Optimum)
 Reaction Catalyzed:

 Principle:
o Catalyze the hydrolysis of various [Amylase – smallest enzyme & produced from acinar cells,
phosphomonoesters at an acid Ph
detected in urine sample]
 Activators:
o Magnesium
o Zinc
 Tissue Sources:
o Prostate [Prostate is the richest tissue source]
o Bone Liver, Spleen, Kidneys, Platelets
 Clinical Significance:
o Prostatic Carcinoma
o Forensic Chemistry [presumptive evidence not
conclusive not definitive]
 aids in the investigation of rape [sample:
vaginal washing; vaginal washing check for
seminal fluid check ACP activity present 4
days]
o Platelet destruction
 Idiopathic Thrombocytopenic Purpura (ITP)
o Bone diseases
 Paget's disease
 Breast CA w/ bone metastasis
 Gaucher's disease
 [Bone, ACP is associated with osteoclasts]
 Prostate Specific Antigen
o more specific than ACP
o Substrate: thymolphthalein monophosphate
o Inhibitor: tartrate

 Assay:
o Thymolphthalein monophosphate
 Substrate for quantitative endpoint
reactions
o Alpha-naphthyl phosphate
 Substrate for continuous monitoring
methods

 Source of Error:
o Serum separated from RBC ASAP
o If not assayed ASAP serum should be frozen
or acidified to a pH < 6.5
AMYLASE (AMS)
 E.C.3.2.1.1
 1,4-D-Glucan Glucanohydrolase

MONTEMOR, DJ 1
2
 Activators: o Colipase and bile salts
o Calcium, Chloride, Iodine, Bromine  Tissue Sources:
 Principle: o Pancreas [Lipase is the most specific]
o Catalyze the breakdown of starch and o Stomach, Small Intestine
glycogen to glucose  Source of Error
 Isoenzymes:
o S-type (salivary) [other name: Ptyalin]
o P-type (pancreatic) [other name: Amylopsin]
 Tissue Sources:
o Salivary glands
o Pancreas
 Source of Error:
o Relatively unaffected by hemolysis
o Contamination of saliva
 Clinical
Significance:
o Acute Pancreatitis
o Salivary gland disorders
 Mumps
 Parotitis
o Intra-abdominal disease
o Hyperamylasemia
o Macroamylasemia
ACUTE PANCREATITIS
RISE 2-12 hours
PEAK 24 hours
NORMALIZE 3-5 days
 Assay:
o Amyloclastic
 measures disappearance of the substrate
 STARCH+ IODINE = bluish black color
 Add SERUM = diminishing bluish black
color
 Decrease in color is proportional
to AMS concentration
o Saccharogenic
 measures the appearance of the product
 Form sugar units  monosaccharide 
glucose
 Concentration of reducing sugars is
proportional to AMS activity
o Chromogenic
 measures color intensity
 STARCH+CHROMOGENIC DYE = insoluble
 dye-substrate complex+ SERUM =
hyrdrolysis
 Increase in color intensity is
proportional to AMS activity
LIPASE (LPS)
 E.C.3.1.1.3
 Triacylglycerol acylhydrolase
 Reaction
Catalyzed:

 Activator:
o Calcium
 Principle:
o Catalyzes the partial hydrolysis of dietary
TAG in the intestine to the 2
monoglyceride and fatty acids
 Accelerators: [enhances the reaction, not require
for the reaction to occur]

MONTEMOR, DJ 1
3
 o Hemolysis
 Clinical Significance:
GLUCOSE-6-PHOSPHATE DEHYDROGENASE
o Acute Pancreatitis
 almost exclusively for this diagnosis (G6PD)
o Intra-abdominal diseases  E.C.1.1.1.49
ACUTE PANCREATITIS  D-Glucose-6- Phosphate" NADP+1- Oxidoreductase
RISE 2-12 hours  Reaction Catalyzed:
PEAK 24 hours
NORMALIZE 3-5 days
 Assay:
o Cherry-Crandall Method
 measures the liberated fatty acids by
titration after a 24-hour incubation
 substrate olive oil
 OLIVE OIL + SERUM = measure FA by
titration after 24-hr incubation
o Modified Cherry-Crandall
 substrate triolein
o Turbidimetry
 principle: as the fats are hydrolyzed by
LPS the particles disperse, and the rate of
clearing can be measured as an
estimation of LPS activity  Principle:
o Coupled Enzyme Reaction o Catalyze the oxidation of G6P to
6- phosphogluconate or lactone
 principle based on coupled reactions with
peroxidase or glycerol kinase o Maintains NADPH in reduced form in
RBCs [Glutathione - protect RBCs from oxidation]
GAMMA-GLUTAMYL TRANSFERASE (GGT)
 Clinical Significance:
 E.C.2.3.2.2
o G-6-PD Deficiency
 (5-Glutamyl) Peptide Amino Acid-5-Glutamyl
 inhented sex-linked trait
Transferase  can cause hemolytic anemia
o Increased G-6-PD
 myocardial infarction
 Principle:  megaloblastic anemia
o Involved in the transfer of the gamma  Tissue Sources:
glutamyl residue from gamma-glutamyl o RBCs, Adrenal Cortex, Spleen, Thymus,
peptides to amino acids, H2O, and other Lymph Nodes, Lactating Mammary Glands
small peptides  Assay:
 Physiology: o Sample: red cell hemolysate or serum
o Involved in peptide and protein synthesis
o Regulation of tissue glutathione levels
o Transport of AA across cell membranes
 Glutathione ANGIOTENSIN-CONVERTING ENZYME (AMS)
o Glutamyl donor  E.C.3.4.15.1
 Tissue Sources:  Angiotensin I-Converting Enzyme/ Kininase 11/
o Liver [marker for chronic alcoholism] Peptidyl- Dipeptidase A
o Kidneys Brain Prostate Pancreas
 Clinical Significance:
o Hepatobiliary disorders
o Patients receiving enzyme inducing drugs
 warfarin, phenobarbital, phenytoin
o Marker for Chronic Alcoholism
o Unknown etiology
Hepatocellular Biliary
involvement obstruction
GGT Normal Increased
ALP Increased Increased
 Assay:
o substrate: gamma-glutamyl-p-nitroanilide
o abs: 405-420nm

 Source of Error:
o Relatively unaffected by hemolysis

MONTEMOR, DJ 1
4
[Renin–Angiotensin–Aldosterone System (RAAS) -
system involves enzymes and hormones to
maintain water and sodium in the body and also a
vasoconstriction] [Angiotensinogen (-ogen =
inactive) - go to kidneys where Renin is found
(enzyme) convert Angiotensin I  Angiotensin II
(potent form). Angiotensin I go to the Lungs where
ACE is found to convert Angiotensin I 
Angiotensin II (potent enough to induced
vasoconstriction) & stimulate the Aldosterone
(Na & Water Retention), & Argenine Vasopressin
(ADH)]
 Principle:
o Hydrolysis of peptide bonds at the
free C-terminus, releasing dipeptide
reaction

MONTEMOR, DJ 1
5
 Tissue Sources:
o Lungs & Testes
 Physiology:
o Converts angiotensin I to angiotensin II in RAAS
o Inactivation of bradykinin in Kallikrein-Kinin
System
o Modulate peripheral vascular resistance and
renal & cardiovascular function  Physiology:
o Predominantly found in he endothelial o Cytoplasmic membrane- bound phosphatase
cell membranes throughout o Acts only on nucleotides
the body o A metalloenzyme (zinc)
o DIAG SIGN: sarcoidosis o Widely distributed in the body, predominantly
 Assay: attached to cell membranes (similarly to ALP
o Principle: ability to cleave synthetic and GGT)
peptides, releasing hippuric acid  Tissue Source:
ACETYL CHOLINESTERASE (ACHE) o LIVER
 E.C. 3.1.1.7  Clinical Significance:
 Acetylcholine acylhydrolase o Commonly used to determine if the source
 Also known as: “True” Cholinesterase of elevated ALP is from liver or bone
 Reaction Catalyzed: o Cholestatic disorders
o Ovarian cancer
o Rheumatoid arthritis
 Assay:
o Made difficult because other phosphatases
are capable of cleaving the substrate
o Uses ALP inhibitors
 Principle: o Chelating agents inhibit activity
o Hydrolyzes acetyl-B- methylcholine
 Physiology:
o Found in synapse of the nerve and RBC Not
normally found in maternal blood and
amniotic fluid
 Clinical Significance:
o Organophosphate poisoning
 decreased levels
o Neural tube defects
 qualitative analysis in amniotic
fluid SampleHemolysate of washed
RBC
PSEUDO-CHOLINESTERASE (PCHE)
 E.C.3.1.1.6
 (5-Glutamyl) Peptide: Amino Acid-5-
Glutamyl Transferase

 Principle:
o Hydrolyzes butyryl- and benzoylcholine
 Sample: serum
 Clinical Significance:
o Organophosphate poisoning
 PChE activity (serum) falls before AChE
activity (RBC)
o Liver disease
 reflects liver synthetic function rather
than hepatocyte injury
 Assay:
o ELLMAN'S METHOD
o principle: released thiocholine react
with dilipbisnitrobenzoic acid
o substrate: acylthiocholine ester
o product: 5-mercapto-2 nitro benzoic acid
5'N NUCLEOTIDASE (5’N)
 EC 3 1 3 5
 5’-Ribonucletide Phosphohydrolase

MONTEMOR, DJ 1
6
 Reaction Catalyzed:

MONTEMOR, DJ 1
7