Other Clinically Important Enzymes • Assays for enzyme activity

1. Acid Phosphatase (3.2.3.1) /01

𝑝– 𝑛𝑖𝑡𝑟𝑜𝑝ℎ𝑒𝑛𝑦𝑙𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒 𝑝– 𝑛𝑖𝑡𝑟𝑜𝑝ℎ𝑒𝑛𝑜𝑙 + 𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
• Systematic name: Orthophosphoric monoester phosphohydrolase (acid optimum)
• Common abbreviation: ACP o Thymolphthalein monophosphate – quantitative end point reaction
• Standard abbreviation: ACP o α-naphthyl phosphate – continuous monitoring method

• Present in lysosomes • Sources of error

o Sac-like organelles containing various hydrolytic enzymes
pH Stability Hemolysis
• Tissue sources: < 6.5 Stable Affected (reject sample)
o Prostate
o Spleen 2. Glucose-6-Phosphate Dehydrogenase (E.C. 1.1.1.49)
o Bone •
+
Systematic name: D-Glucose-6- phosphate:NADP 1-oxidoreductase
o RBCs
• Common abbreviation: G-6PDH
o Platelets
• Standard abbreviation: G-6-PD
• Reference value
• Predominantly a dimer; tetramer (pH dependent)
o 0 to 3.5 ng/mL
• Composed of 500 subunits
/01
𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑚𝑜𝑛𝑜𝑒𝑠𝑡𝑒𝑟 + 𝐻- 𝑂 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 + 𝑃𝑂6 7 • Tissue sources:
o pH: 5 o Adrenal cortex
o Spleen
• ACP determining genes o Thymus
o Erythrocytic ACP o Lymph nodes
§ Chr. 2 – polymorphic o Lactating mammary gland
§ Chr. 19 – encodes TR-ACP (tartrate ACP) expressed in osteoclasts o RBC
o Lysosomal & prostatic ACP – tartrate-inhibited ACP
§ Chr. 11 • Reference value:
§ Chr. 13 o 8-14 U/g Hgb

• Diagnostic significance • Assay for enzyme activity

o Tartrate-inhibited ACP @A1B
§ Carcinoma of prostate 𝐺6𝑃 + 𝑁𝐴𝐷𝑃 6– 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑔𝑙𝑢𝑐𝑜𝑛𝑎𝑡𝑒 + 𝑁𝐴𝐷𝑃𝐻 + 𝑁𝐻 E
o TR-ACP
• Diagnostic significance
§ Bone involvement:
o If value exceeds 18 U/g Hgb: hemolytic anemia
• Paget’s disease
o Gene for G6PD located at X chromosome
• Hyperparathyroidism with skeletal involvement
§ Presence of malignant invasion of bones by cancer
o Class of severity of G6PD deficiency
§ Non-bone involvement:
Class I Severe anemia associated with chronic hemolytic anemia
• Gaucher disease
Class II Severe deficiency usually without hemolytic anemia
< 10% residual activity
o PSA – prostate-specific antigen
§ Replaced ACP Class III Moderate to mile deficiency
10-60% residual activity
o Assays useful in forensic clinical chemistry
§ Investigation of rape: vaginal washings tested for seminal ACP activity Class IV Very mild or no deficiency
Class V Increased activity
 o Clinical syndromes associated with G6PD deficiency • Other sources:
§ Drug-induced hemolysis o Semen, testes
§ Favism o Ovaries, fallopian tubes
• Caused by inherited G6PD deficiency o Striated muscle, lungs, adipose tissue, colostrum
• Often observed in males o Tears, milk
• Develop hemolytic anemia on consumption of beans or pollen of
beans • Reference value:
§ Infection-induced hemolysis o Serum: 25-130 U/L
§ Neonatal jaundice o Urine: 1-15 U/h
§ Chronic non-spherocytic hemolytic anemia
• Diagnostic significance
Pancreatic Enzymes Acute pancreatitis 5-8h: elevated
1. Amylase (E.C. 3.2.1.1) 12-72h: 4-6 x URL
rd th
• Systematic name: 1,4-D-Glucan glucanohydrolase 3 -4 day: returns to normal
• Common abbreviation: AMS Biliary tract diseases 4 x URL
• Standard abbreviation: AMY Intraabdominal events 4 x URL
Peritonitis & acute appendicitis 2-3 x URL
• Hydrolyze starch and glycogen Renal impairment No more than 5 x URL
o Important in physiologic digestion of starches Hyperamylasemia 50 x URL
• Calcium metalloenzyme Salivary gland lesions 4 x URL
2+
o Ca required for functional integrity Alcoholic intoxication 3 x URL
• Activators: Cl, Br, cholate, monohydrogen monophosphate, nitrate Macroamylasemia 2-3 x URL
o Cl, Br: most effective activators
o Macroamylasemia can cause hyperamylasemia
• Digestion of starch begins in mouth with the hydrolytic action of AMS § Not filtered to glomeruli due to size: > 200 kDa
o Activity is very short: intestinal / gastric contents inactivate AMS activity
o Pancreatic AMS performs major digestive action • Assay for enzyme activity
• Smallest enzyme: normally filtered by glomerulus because of size a. Amyloclastic
• Measures the disappearance of starch substrate
• Isoenzymes – products of 2 closely linked loci in chr. 1 • Uses iodine to produce starch-iodine complex (dark blue color)
a. S-type (salivary) • AMS allowed to act on starch substrate
• Initiates hydrolysis of starch b. Saccharogenic
b. P-type (pancreatic) • Classic reference method
• Synthesized by acinar cells • Measures the appearance of the product
• Secreted to the intestines • Reported in Somogyi units
• Starch o Number of mg of glucose released in 30 minutes at 37°C
Amylose Amylopectin c. Chromogenic
• Long, unbranched chain of glucose • Branched chain polysaccharide • Measures the increasing color from production of product coupled with a
molecules • 1-6 linkages at the branch points chromogenic dye
• Linked by 1,4-glycosidic bonds d. Continuous monitoring
o α-AMS attacks glycosidic bonds to • Coupling of several enzyme systems to monitor amylase activity
produce degradation products: • Absorbance: 340 nm
glucose, maltose, dextrins /FG
𝑚𝑎𝑙𝑡𝑜𝑝𝑒𝑛𝑡𝑜𝑠𝑒 𝑚𝑎𝑙𝑡𝑜𝑡𝑟𝑖𝑜𝑠𝑒 + 𝑚𝑎𝑙𝑡𝑜𝑠𝑒
(intermediate chains) H7IJKLMNOPQNR
𝑚𝑎𝑙𝑡𝑜𝑡𝑟𝑖𝑜𝑠𝑒 + 𝑚𝑎𝑙𝑡𝑜𝑠𝑒 5– 𝑔𝑙𝑢𝑐𝑜𝑠𝑒
URVMWOXQNR
• Tissue sources: 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 + 𝐴𝑇𝑃 𝐺6𝑃 + 𝐴𝐷𝑃
o Acinar cells of pancreas @A1B
𝐺6𝑃 + 𝑁𝐴𝐷 E 5, 6– ℎ𝑜𝑠𝑝ℎ𝑜𝑔𝑙𝑢𝑐𝑜𝑛𝑜𝑙𝑎𝑐𝑡𝑜𝑛𝑒 + 5 𝑁𝐴𝐷𝐻
o Salivary glands
 • Sources of error • Modified Cherry Grandall
Stability o Used triolein instead of olive oil
Sample
Temperature Time b. Turbidimetric method
RT 1 day Administration of morphine and other • More simple, more rapid
4°C 2 hours opiates: falsely elevated serum AMS • Uses olive oil / oleic acid with simultaneous decrease in turbidity of
levels mixture reaction
• Absorbance: 340 nm
2. Triacylglycerol lipase (E.C. 3.1.1.3) c. Colorimetric method
• Systematic name: Triacylglycerol acylhydrolase • Coupled reactions with enzymes:
• Common abbreviation: LPS o Peroxidase
• Standard abbreviation: LPS o Glycerol kinase
• Absorbance: 580 nm
• LPS – single chain lipoprotein • End product: 4-methyl-resorufin ester
• MW: 48 kDa
• Isometric point (pI): 5.8 1, 2– 𝑂– 𝑑𝑖𝑙𝑎𝑢𝑟𝑦𝑙– 𝑟𝑎𝑐– 𝑔𝑙𝑦𝑐𝑒𝑟𝑜– 3– 𝑔𝑙𝑢𝑡𝑎𝑟𝑖𝑐 𝑎𝑐𝑖𝑑– 4– 𝑚𝑒𝑡ℎ𝑦𝑙– 𝑟𝑒𝑠𝑜𝑟𝑢𝑓𝑖𝑛 𝑒𝑠𝑡𝑒𝑟
[1G Eab – NdMXeQXRMKN, ab f
• Gene: chr. 10 𝑡𝑟𝑖𝑐𝑎𝑟𝑏𝑜𝑛𝑖𝑐 𝑎𝑐𝑖𝑑 𝑒𝑠𝑡𝑒𝑟 𝑔𝑙𝑢𝑡𝑎𝑟𝑎𝑡𝑒
• Requires bile salts + colipase + 𝑚𝑒𝑡ℎ𝑦𝑙 𝑟𝑒𝑠𝑜𝑟𝑢𝑓𝑖𝑛 (𝑟𝑒𝑑)
• Small molecule: totally reabsorbed by renal tubules à not normal in urine
• Sources of error
• Tissue source: pancreas Stability
Hemolysis
• Other sources: stomach, small intestines Temperature Time
RT 1 week Inhibits the activity of serum
• Reference value: 4°C 3 weeks LPS: falsely low values
o 0-1.0 U/mL
3. Trypsin (E.C. 3.4.21.4)
[1G
𝑡𝑟𝑖𝑎𝑐𝑦𝑙𝑔𝑙𝑦𝑐𝑒𝑟𝑜𝑙 + 2 𝐻- 𝑂 2– 𝑚𝑜𝑛𝑜𝑔𝑙𝑦𝑐𝑒𝑟𝑖𝑑𝑒 + 2 𝑓𝑎𝑡𝑡𝑦 𝑎𝑐𝑖𝑑𝑠 • Systematic name: Trypsin
o pH: 8.6-9.0 • Common abbreviation: TRY
• Standard abbreviation: TRY
• Diagnostic significance
Acute pancreatitis 2-50x URL • Serine protease – hydrolyzes peptide bonds
Biliary tract diseases Elevated • Inhibitors:
Obstruction of pancreatic duct Elevated o α1-antitrypsin
Reduced glomerular filtration rate Elevated o α2-macroglobulin

o Acute pancreatitis • Proenzyme / zymogen

§ 4-8h: increase
§ 24h: peak Trypsinogen-1 Trypsinogen-2
§ 8-14d: decreases Concentration Fourfold
Mobility Cationic Anionic
• Methods of analysis Molecular weight 25.8 kDa 22.9 kDa
a. Titrimetric method pI values 4.6 – 6.5 > 6.5
• Reference procedure
• Uses olive oil / oleic acid titrated with dilute alkali • Stimulated by:
• Cherry Grandall method (classic method) o Vagus nerve
o Olive oil – substrate o Cholecystokinin pancreozymine (intestinal hormone)
o Phenolphthalein – indicator • Secreted in duodenum
o 0.5N NaOH – titrant
 • Diagnostic significance:
o Trypsinogen – major form found in serum of healthy individuals • Amino acids to differentiate from chymotrypsin
Acute pancreatitis 2-400 x URL o Ala, Val, Leu
Renal failure Elevated
Chronic pancreatitis without steatorrhea Plasma concentration does not differ from • MW: 26 kDa
those of healthy individuals • Synthesized by the acinar cells
Chronic pancreatitis with steatorrhea Extremely low o As a preproelastase to proelastase
Relapsing phase of chronic pancreatitis Elevated o Stored in zymogen granules
Carcinoma of the pancreas High, normal, low o Activated to elastase by TRY in the duodenum
Cystic fibrosis High in neonates
Falls as disease progresses • Measurement
o Stool: most reliable and sensitive
o Acute pancreatitis – trypsinogen-1 rises in parallel with serum amylase level, § Enzyme stable in stool for 1 week
more difficult to perform (requires several hours to complete procedure) o Diagnosis of chronic pancreatic insufficiency
§ Provides no information helpful to the therapeutic management
4. Chymotrypsin (E.C. 3.4.21.1) o LRL: 200 µg/g stool
• Systematic name: Chymotrypsin
• Common abbreviation: CHY
• Standard abbreviation: CHY

• Serine protease
• Reference value:
o 12 U/g stool
• Amino acids to differentiate from elastase
o Trp, Leu, Tyr, Phe

• Zymogens
o Chymotrypsinogen-1
o Chymotrypsinogen-2

• MW: 25 kDa
• Bound in plasma by:
o α1-antitrypsin
o α1-macroglobulin
• Activity in stool: constant at RT for 7 days
• Trypsin – converts chymotrypsinogen to chymotrypsin
• Chymotrypsin: more resistant than trypsin, enzyme of choice in stool assay

• Diagnostic significance
o Chronic pancreatic insufficiency – below LRL

5. Elastase-1 (E.C. 3.4.21.36)

• Systematic name: Elastase
• Common abbreviation: E1
• Standard abbreviation: E1

• A carboxyendopeptidase that hydrolyzes the native elastin (major structural

fibrous protein in connective tissues) with special affinity for the carboxyl group
 Liver Enzymes
1. 5’ Nucleotidase (E.C. 3.1.3.5) o LD6 – alcohol dehydrogenase
• Systematic name: 5’-Ribonucleotide phosphohydrolase § Present in patients with arteriosclerotic cardiovascular failure
• Common abbreviation: NTP § May reflect liver injuries related to severe circulatory deficiency
• Standard abbreviation: NTP § Identified in sera of very sick patients

• Glycoprotein localized in cytoplasmic membrane Isoenzyme Tissue Disorder

• Phosphatase that acts on adenosine-5’ monophosphate to yield adenosine LD1 Heart Myocardial infarction
XKLJRMeOPQNR RBCs Hemolytic anemia
o 𝑎𝑑𝑒𝑛𝑜𝑠𝑖𝑛𝑒– 5i – 𝑚𝑜𝑛𝑜𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒 𝑎𝑑𝑒𝑛𝑜𝑠𝑖𝑛𝑒
LD2 Heart Megaloblastic anemia
§ pH: 6-6.7
RBCs Acute renal infarct
Hemolyzed specimen
• Tissue source: liver
LD3 Lung Pulmonary embolism
• Reference value: 3-9 U/L (37°C)
++ Lymphocytes Extensive pulmonary pneumonia
• Activator: Zn
Spleen Lymphocytosis
Pancreas Acute pancreatitis
• Diagnostic significance:
Carcinoma
Hepatobiliary disorder 3-6 x URL
LD4 Liver Hepatic injury or inflammation
Infectious hepatitis Moderately elevated
LD5 Skeletal muscle Skeletal muscle injury
Ovarian carcinoma Elevated
Rheumatoid arthritis Elevated o Normal: LD2 > LD1
§ LD flipped pattern – LD1 > LD2
2. Lactate Dehydrogenase (E.C. 1.1.1.27) • Indicative of acute MI
+
• Systematic name: L-Lactate: NAD oxidoreductase • Not specific to cardiac diseases
• Common abbreviation: LD
• Standard abbreviation: LD Isoenzyme %
LD1 14-26
• Tissue sources: LD2 29-39
o Heart, liver, skeletal muscles, kidney, RBCs
LD3 20-26
• Other sources:
LD4 8-16
o Lungs, smooth muscle, brain
LD5 6-16
• Reference value: 100-225 U/L (37°C)

[B • Diagnostic significance
• 𝐿– 𝑙𝑎𝑐𝑡𝑎𝑡𝑒 + 𝑁𝐴𝐷 E 𝑝𝑦𝑟𝑢𝑣𝑎𝑡𝑒 + 𝑁𝐴𝐷𝐻 + 𝐻 E
o pH: Acute myocardial infarction
§ Forward: 8.8-9.8 Pulmonary embolism
§ Reverse: 7.4-7.8 Slightly elevated
+ + Viral hepatitis
o NAD – H acceptor 2-3 x ULN
Cirrhosis
o Strongly favors reduction of pyruvate to lactate
Infectious mononucleosis
§ P à L: 3-4x faster
Pernicious anemia
Highest level
Hemolytic anemia
• Subunit compositions of 5 isoenzymes of LD in order of decreasing anodal mobility
Skeletal muscle disorders and some leukemias Some are elevated
in alkaline medium
Acute lymphoblastic leukemia Marked increase
LD1 HHHH H4
LD2 HHHM H3M
LD3 HHMM H2M2
LD4 HMMM HM3
LD5 MMMM M4
3. Cholinesterase • Method of analysis

Acylcholinesterase Acylcholine acylhydrolase NRnKo 0Ul

𝐵𝑢𝑡𝑦𝑟𝑦𝑙 𝑡ℎ𝑖𝑜𝑐ℎ𝑜𝑙𝑖𝑛𝑒 𝑎𝑐𝑒𝑡𝑦𝑙 𝑡ℎ𝑖𝑜𝑙𝑖𝑛𝑒 𝑒𝑠𝑡𝑒𝑟 𝑏𝑢𝑡𝑦𝑟𝑎𝑡𝑒 + 𝑡ℎ𝑖𝑜𝑐ℎ𝑜𝑙𝑖𝑛𝑒
E.C 3.1.1.7 3.1.1.8
Systematic name Acetylcholine Acylcholine acylhydrolase, 𝑡ℎ𝑖𝑜𝑐ℎ𝑜𝑙𝑖𝑛𝑒 + 𝐷𝑇𝑁𝐵 𝑐𝑜𝑙𝑜𝑟𝑙𝑒𝑠𝑠 ↔ 𝑚𝑖𝑥𝑒𝑑 𝑑𝑖𝑠𝑢𝑙𝑓𝑖𝑑𝑒 + 5 𝑀𝑁𝐵𝐴 (𝑐𝑜𝑙𝑜𝑟𝑒𝑑)
acetylhydrolase ChE
Other names True cholinesterase Pseudo cholinesterase o DTNB – disulfide agents: 5,5’-dithiobis(2-nitrobenzoate)
Choline esterase I Choline esterase II o MNBA – 5-mercapto-2-nitrobenzoic acid
Serum cholinesterase
Butyryl cholinesterase o Absorbance: 410 nm
Location Erythrocytes, lungs, spleen, Liver, pancreas, heart, white
nerve endings, gray matter matter of brain, serum 4. Glutamate dehydrogenase (E.C. 1.4.1.3)
of brain • Systematic name: L-glutamate:NAD(P) oxidoreductase, deaminase
Acetyl-β-methyl choline Hydrolyzes (RBC) Does not • Common abbreviation: GLD
Benzoyl choline Does not Hydrolyzes (Serum) • Standard abbreviation: GLD

• Responsible for the prompt hydrolysis of acetylcholine released at the nerve • Tissue sources: liver, heart muscle, kidney
endings to mediate transmission of the neural impulse across the synapse • Other sources: brain, skeletal muscle, leukocytes
o Choline
§ Water-soluble vitamin • Diagnostic significance: hepatocellular damage
§ Basic compound formed in various foods (i.e. egg yolk, legumes) • Reference values:
§ Synthesized in liver, component of lecithin o M: 8 U/L (37°C)
• Lecithin – precursor of acetylcholine o F: 6 U/L (37°C)
0Ul @[B
• 𝑎𝑐𝑒𝑡𝑦𝑙𝑐ℎ𝑜𝑙𝑖𝑛𝑒 𝑏𝑟𝑜𝑚𝑖𝑑𝑒 𝑎𝑐𝑒𝑡𝑎𝑡𝑒 + 𝑐ℎ𝑜𝑙𝑖𝑛𝑒 𝑏𝑟𝑜𝑚𝑖𝑑𝑒 𝐿– 𝑔𝑙𝑢𝑡𝑎𝑚𝑎𝑡𝑒 𝑘𝑒𝑡𝑜𝑚𝑖𝑛𝑜 𝑎𝑐𝑖𝑑 + 2– 𝑜𝑥𝑜𝑔𝑙𝑢𝑡𝑎𝑟𝑎𝑡𝑒

• Diagnostic significance
a. Liver function: sensitive indicator of synthetic capacity of liver Sources of Error
b. Insecticide poisoning
c. Detection of patients with atypical forms of enzymes Stability
• At risk for prolonged responses to certain muscle relaxants used in Enzyme Hemolysis
Temperature (°C) Time
surgical procedures AST 2-8 3-4 d Increased concentration
ALT 4 3-4 d Does not interfere
Acute hepatitis 30-50% decrease 4 No loss of activity Does not interfere
Cirrhosis 50-70% decrease GGT
for 1 week
Carcinoma with metastases 25 Inc. approx. 3-10% Does not interfere
Obstructive jaundice Normal except when malignant ALP on standing
4 Several hours
• Reference values: 25 48 h Unacceptable sample
o M: 40-78 U/L (37°C) LDH
4 (LD5) 24 h Several hours
o F: 33-76 U/L (37°C) 4 4d
o Heterozygous genotype: 5-35 U/L (37°C) NTP
-20 4 mos
o Atypical homozygotes: 1-4 U/L (37°C) 2-8 Several weeks
o At birth: 50% lower than adult values ChE
-20 Several years
o 3-6 years old: exceed adult value by 30%
4 48 h
o Pregnancy: significant decrease (-30%) GLD
-20 Several weeks
Muscle Enzymes
1. Creatine kinase (E.C. 2.7.3.2) o CK-Mi/Mt (Mitochondrial CK)
• Systematic name: ATP:creatine N-phosphotransferase § Found at chr. 15
• Common abbreviation: CPK § Bound to the exterior surface of the inner mitochondrial membrane of
• Standard abbreviation: CK muscles, brain and liver
§ Cathodal to CK-MM
• Dimer § Not present in normal serum
• MW: 82 kDa • Only present if tissue damage is extensive, releasing mitochondria
• Catalyzes reversible phosphorylation of ATP to ADP and cell wall
§ Incidence: 0.8-1.7%
0u, FXvw § Detected in cases of malignant tumor and cardiac abnormalities
𝐶𝑟𝑒𝑎𝑡𝑖𝑛𝑒 + 𝐴𝑇𝑃 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑐𝑟𝑒𝑎𝑡𝑖𝑛𝑒 𝑐𝑟𝑒𝑎𝑡𝑖𝑛𝑒 𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒 + 𝐴𝐷𝑃
• Electrophoresis
o pH:
§ Forward: 9
Cathode Anode
§ Reverse: 6.7 MI x MM Macro MB BB
(–) (+)
o Phosphocreatine – phosphorylation reservoir
• Diagnostic significance
• ATP à ADP when muscle contracts
2+
• Activating ion: Mn
Duchenne sex-linked muscular dystrophy Infancy & childhood: highest level
o Excess Mg inhibits reaction
As patient gets older: falls
Asymptomatic female: 50-80% w/ 3-6x URL
• Tissue sources: skeletal muscles, heart muscles, brain tissue
Muscular dystrophy (all types) 50-100x URL
• Other sources: bladder, placenta, GIT, thyroid, uterus, kidney, lung, prostate,
Neurologic muscular diseases Normal
spleen, liver, pancreas
Malignant hyperthermia Elevated
Uremic myopathy Elevated (CK-MB)
• Reference values:
Acute rhabdomyolysis 200x URL
o Total CK:
§ M: 15-160 U/L (37°C) Hypothyroid activity Inverse relp: 5x URL (CK-MM)
§ F: 15-130 U/L (37°C) Maternal total serum Childbirth: 6x URL (CK-BB)
o CK-MB: < 6% total CK Myocardial damage 20% of all CK-MB
Begins to rise within 4-8 hours
• 2 subunits: products of chr. 14 & 19 Peaks at 12-24 hours
o B&M Returns to normal within 48-72 hours
Acute cerebral disorders (CK-MB) – not normally expected
• Isoenzymes: found in the cytosol of cell Lung cancer
o BB (CK-1) – brain
o MB (CK-2) – hybrid o AMI – consequent increase of LDH (period of 48 hours)
o MM (CK-3) – muscle o
o Macro-CK • Troponin 1 & Troponin T – non-enzyme proteins used as a more sensitive specific
§ Largely comprised with BB marker for myocardial damage
§ Often transiently found in serum of up to 6% of hospitalized patients
§ Complexed with IgG, sometimes IgA • Assay for enzyme activity
Form Type 1 Type 2 o Forward reaction:
Characteristic Complex of CK Oligometric CK-Mt § Coupled with PK-LD-NADH system
Prevalence 0.8 & 2.3% 0.5 & 2.6%
0u
Occurrence Women (>50 years old) Adults: with malignancies or liver disease 𝑐𝑟𝑒𝑎𝑡𝑖𝑚𝑒 + 𝐴𝑇𝑃 𝑐𝑟𝑒𝑎𝑡𝑖𝑛𝑒 𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒 + 𝐴𝐷𝑃
Children: notable in tissue distress 1u
𝐴𝐷𝑃 + 𝑃ℎ𝑜𝑠𝑝ℎ𝑜𝑒𝑛𝑜𝑙𝑝𝑦𝑟𝑢𝑣𝑎𝑡𝑒 𝑝𝑦𝑟𝑣𝑎𝑡𝑒 + 𝐴𝑇𝑃
 [B
𝑝𝑦𝑟𝑢𝑣𝑎𝑡𝑒 + 𝑁𝐴𝐷𝐻 + 𝐻 E 𝑙𝑎𝑐𝑡𝑎𝑡𝑒 + 𝑁𝐴𝐷 E
§ No reverse reaction without CK
§ pH: 9.0
§ Absorbance: 340 nm

o Reverse reaction
§ Coupled with hexokinase-G6PD-NADP system

0u
𝑐𝑟𝑒𝑎𝑡𝑖𝑛𝑒 𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒 + 𝐴𝐷𝑃 𝑐𝑟𝑒𝑎𝑡𝑖𝑛𝑒 𝐴𝐷𝑃
bu
𝐴𝑇𝑃 + 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝐴𝐷𝑃 + 𝑔𝑙𝑢𝑐𝑜𝑠𝑒– 6– 𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
@A1B
𝐺6𝑃 + 𝑁𝐴𝐷𝑃𝐻 E 6– 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑔𝑙𝑢𝑐𝑜𝑛𝑎𝑡𝑒 + 𝑁𝐴𝐷𝑃𝐻

§ pH: 6.8
§ 2-6x faster than forward reaction
§ Proposed by Oliver, modified by Rosalki
§ Common method used

• Sources of error

Stability
Hemolysis
Temperature Time
Stored in dark at 4°C 7 days Elevated CK activity
–4°C 1 month*
* Treated with sulfhydryl activator