[CC-LAB] Lactate Dehydrogenase and Creatine Kinase

Lactate + NAD ⇌ Pyruvate + NADH

Reaction for Lactate Dehydrogenase

Lactate Dehydrogenase

It is a zinc-containing enzyme that is part of the glycolytic pathway and Krebs cycle.

Four subunits of two possible forms .

(H and M)

Tetrameric molecule of Lactate Dehydrogenase contains ____.

Lactate Dehydrogenase

Referred to as tissue non-specific marker of inflammation.

100 - 255 u/L

Reference Range for Forward Reaction in Lactate Dehydrogenase

[Good to know lang]

80 - 280 u/L

Reference Range for Reverse Reaction in Lactate Dehydrogenase

[Good to know lang]

LD1

LD2

LD3

LD4

LD5

Isoenzymes of Lactate Dehydrogenase

LD1

Fastest Isoenzyme towards Anode of Lactate Dehydrogenase
(Most Negatively Charged)

LD5 > LD4 > LD3 > LD2 > LD1

Migration of Lactate Dehydrogenase Isoenzymes

(Positive to Negatively Charged)

LD2

Highest Concentration in Serum of Lactate Dehydrogenase Isoenzymes

LD2 > LD1 > LD3 > LD4 > LD5

Concentration Levels in Serum of Lactate Dehydrogenase Isoenzymes

LD4 and LD5

Lactate Dehydrogenase Isoenzymes that are cold-labile

Alpha-hydroxybutyric Dehydrogenase (HBD)

Has the same kinetic properties as LD1, but does not act on the substrate lactate.

The substrate used is ketobutyrate instead of lactate

Main difference between Alpha-hydroxybutyric Dehydrogenase and LD1

Rosalki-Wilkinson Method

Method used in measuring Alpha-hydroxybutyric Dehydrogenase

Ketobutyrate + NAD ⇌ Pyruvate + NADH

Reaction for Alpha-hydroxybutyric Dehydrogenase

LD6

Lactate Dehydrogenase Isoenzyme that denotes the presence of the alcohol dehydrogenase enzyme.

Drug Hepatotoxicity

Obstructive Jaundice

Alcohol dehydrogenase is the 6th band in electrophoresis, usually elevated in cases of ____ and ___.

ADD INFO: Another form of LD composed of four C subunits is found in spermatozoa and in semen b

ADD INFO: Another form of LD composed of four C subunits is found in spermatozoa and in semen b

REMEMBER:

REMEMBER:

HHHH

Chain Composition of LD1

Heart

Brain

Erythrocytes

Tissue Rich in LD1

HHHM

Chain Composition of LD2

Heart

Brain

Erythrocytes

Tissue Rich in LD2

HHMM

Chain Composition of LD3

Brain

Kidney

Tissue Rich in LD3

HMMM

Chain Composition of LD4

Liver

Skeletal Muscle

Kidney

Tissue Rich in LD4

MMMM

Chain Composition of LD5

Liver

Skeletal Muscle

Ileum

Tissue Rich in LD5

Pernicious Anemia or Megaloblastic Anemia

Pronounced elevation of Lactate Dehydrogenase is observed in ____.

Myoglobin

Troponin I

Creatine Kinase

Aspartate Transferase

Lactate Dehydrogenase

Cardiac Markers (My TropiCAL)

12 to 24 hours

Lactate Dehydrogenase increase at ____ after the onset of AMI.

48 to 72 hours

Lactate Dehydrogenase peaks at ____ after the onset of AMI.

10 to 14 days

Lactate Dehydrogenase returns to normal at ___.

LD Flip Pattern

The concentration of LD1 increases compared to LD2.

Megaloblastic Anemia

Widespread Carcinomatosis (Metastases)

Systemic Shock and Hypoxia

Hepatitis

Renal Infarction

Conditions with Pronounced Elevation (5 or more times normal)

Myocardial Infarction

Pulmonary Infarction

Hemolytic Conditions

Leukemia

Infectious Mononucleosis

Delirium tremens

Muscular Dystrophy

Conditions with Moderate Elevation (3 to 5 times normal)

Most Liver Diseases

Nephrotic Syndrome

Hypothyroidism

Cholangitis

Conditions with Slight Elevation (Up to 3 times normal)

REMEMBER: Serum is preferred for Lactate Dehydrogenase

REMEMBER: Serum is preferred for Lactate Dehydrogenase

Wacker Method (Forward Reaction)

Measures enzymatic activity as lactate is converted to pyruvate

340 nm

In Kinetic for Lactate Dehydrogenase:

Measures the increase in absorbance at ___ nm as NAD is converted to NADH.

Phenazine Methosulfate

Nitroblue Tetrazolium

In Colorimetric for Lactate Dehydrogenase:

Add ____ and ____.

Phenylhydrazone

In Colorimetric for Lactate Dehydrogenase:

Add Phenazine Methosulfate and Nitroblue Tetrazolium to produce ____.

440 nm or 525 nm.

In Colorimetric for Lactate Dehydrogenase:

Add Phenazine Methosulfate and Nitroblue Tetrazolium producing Phenylhdrazone measured at ____ or ___ nm.

Wrobleuski-La Due (Reverse Reaction)

Measures enzymatic activity as pyruvate is converted to lactate

REMEMBER: Wroblueski-La Due is 3 times faster than Wacker

REMEMBER: Wroblueski-La Due is 3 times faster than Wacker

Creatine Kinase

Catalyzes the transfer of phosphate groups from creatine to form creatine phosphate.

Creatine + ATP ⇌ Creatine Phosphate + ADP

Reaction for Creatine Kinase

Magnesium

Thiol Surface

Creatine Kinase requires ___ and ___.

Zinc

Manganese

Creatine Kinase is inhibited by ___ and ___.

Dimeric

(M and B)

Creatine Kinase is a ___ molecule and composed of two different monomers.

Brain

Skeletal Muscle

Cardiac Muscle

Major Tissue Sources of Creatine Kinase

CK-BB (Brain Type)

CK-BM (Hybrid Type)

CK-MM (Muscle Type)

Isoenzymes for Creatine Kinase

95%

____ of total CK activity is derived from CK-MM

2 to 3 hours

Half-life of CK-BB

15 hours

Half-life of CK-MM

12 hours

Half-life of CK-MB

Mitochondrial CK

CK found within the mitochondria of the cell. It does not appear in the circulation unless more severe cellular damage takes place.

CK-BB

Fastest Isoenzyme of Creatine Kinase

Intramitochondrial CK

It is also known as the MtCK or non-muscle/ubiquitous mitochondrial CK.

Sarcomeric Muscle CK

Acts to support structure and contractility of muscle.

4 to 8 hours

Creatine Kinase-MB increase at ____ after the onset of AMI.

12 to 24 hours

Creatine Kinase peaks at ____.

48 to 72 hours

Creatine Kinase returns to normal at ____.

Duchenne’s Muscular Dystrophy

Highest elevation of CK is found in ___.

REMEMBER: CK is not present in Liver

REMEMBER: CK is not present in Liver

Duchenne’s Muscular Dystrophy

Polymyositis

Dermatomyositis

Myocardial Infarction

[Creatine Kinase]

Conditions with Pronounced Elevation (5 or more times normal)

Severe Exercise, Trauma, Surgical Procedure

Intramuscular Injection

Delirium Tremens

Alcoholic Myopathy

Severe Ischemic Injury

Pulmonary Infarction

Pulmonary Edema

Hypothyroidism

Acute Agitated Psychoses

[Creatine Kinase]

Conditions with Mild or Moderate Elevation (2 to 4 times normal)

ADD INFO: Aldolase is progressive muscular dystrophy, CK is Duchenne’s muscular dystrophy.

ADD INFO: Aldolase is progressive muscular dystrophy, CK is Duchenne’s muscular dystrophy.

Increases

Hemolysis falsely ___ Creatine Kinase.

Adenylate Kinase

____ catalyzes a similar reaction as that of Creatine Kinase.

Adenosine Monophosphate

Binds adenylate kinase

Tanzer-Gilbarg

Forward reaction for Creatine Kinase

Oliver-Rosalki or Rosalki-Hess

Reverse reaction for Creatine Kinase