Fundamentals of Blood Biochemistry

(BCH 220)

DR. MANSOUR GATASHEH

Biochemistry Department, Science College
King Saud University
Class 14:

Plasma lipoproteins and

enzymes

T
Objectives for this lecture

 Discuss the different type of Plasma lipoproteins and

enzymes.

 understand the effect of liver and kidney disease on

the level of plasma lipoproteins and enzymes.
 Plasma Enzymes

Plasma derived Cell Derived:These enzymes

have a high activity in cells &
overflow into the plasma.

These enzymes act on

substrates in plasma & Secretary
their activity is higher in enzymes
plasma than cells. E.g.
coagulation enzymes. Metabolic
enzymes
 Blood plasma contains many enzymes
which are classified into:

1. Functional plasma enzymes

2. Non functional plasma enzyme

 Differences between functional and
non functional enzymes
Functional plasma Non functional plasma
enzymes enzymes
Concentration in Present in plasma in higher Normally, Present in plasma in
concentrations in comparison to very low concentrations in
plasma tissue comparison to tissue

Function Have known functions No known functions

Substrate Their substrates are always Their substrates are absent from
present in plasma plasma

Site of synthesis liver Different organs .g. liver heart,

skeletal muscles and brain

Effect of disease Decrease in liver disease Increase in different organ

diseases

Examples Clotting factors e.g. Prothrombin ALT, AST, CK, LDH, alkaline
Lipoprotein lipase, phosphatase, acid phosphatase
Pseudocholinesterase and lipase
 Small amounts of intracellular
enzymes are present in the
blood as a result of normal cell
turnover.
 'Normal' plasma enzyme levels
reflect the balance between the
rate of synthesis and release
into plasma during cell
turnover, and the rate of
clearance from the circulation.
 The presence of elevated
enzyme activity in the plasma
may indicate tissue damage
that is accompanied by
increased release of
intracellular enzymes
 Source of non functional enzymes
 Cell damage with the release of its content of enzymes
into blood e.g. Myocardial infarction and viral hepatitis
 Obstruction of normal pathways e.g. Obstruction of
bile duct increases alkaline phosphatase
 Increase of the enzyme synthesis e.g. bilirubin
increases the rate of synthesis of alkaline phosphatase
in obstructive liver disease
 Increased permeability of cell membrane as in hypoxia
Medical importance of non
functional enzymes
 Measurement of non functional enzymes is
important for:
1. Diagnosis of diseases as disease of different
organs cause elevation of different plasma
enzymes
2. Prognosis of the disease we can follow up of
the treatment by measuring plasma enzymes
before and after treatment
Disadvantages of enzyme assays

 A major disadvantage for the diagnosis of tissue

damage is their lack of specificity to a particular
tissue or cell type. Many enzymes are common to
more than one tissue.
 This problem may be obviated in 2 ways:
 First,different tissues may contain (and thus release
when they are damaged) two or more enzymes in
different proportions
 Second, some enzymes exist in different forms
(isoforms)
 Isoenzymes
 Isoenzymes (or isozymes) are a group of enzymes that catalyze
the same reaction but they differ in amino acid sequence
 Isoenzymes can be:
– produced by different genes (= true isozymes)
– produced by different posttranslational modification (=
isoforms)

 found in different compartments of a cell

 found in different tissues of an organism
 can be oligomers of various subunits (monomers)
 Isoenzymes
 They differ in:
– electrophoretic mobility
– enzymatic properties
– physical properties (e.g heat stability)
– biochemical properties such as amino acid
composition, immunological reactivities
 Because isoenzymes are originated from
different tissues, their determination give more
information than measurement of total enzyme
activity in plasma
Abnormal plasma enzyme activities

 Aspartate Transaminase (AST)/

Serum Glutamate oxaloacetate (SGOT)
 Diagnostic Significance
 The clinical use of AST is limited mainly to the
evaluation of hepatocellular disorders and skeletal
muscle involvement.
 Post AMI (Acute Myocardial Infarction )
– Rises 6 – 8 hours
– Peaks at 24 hours
– Returns to normal by day 5
 AST levels are highest in acute hepatocellular
disorders, viral hepatitis, cirrhosis.
– Viral hepatitis may reach 100 x ULN (Upper limit of
Normal)
 Diagnostic Significance
 There are two isoenzyme fractions located in
the cell cytoplasm and mitochondria,
– the cytoplasmic isoenzyme is predominant in
serum
– while the mitochondrial one may be increased
following cell necrosis.
 Isoenzyme analysis of AST is not routinely
performed in the clinical laboratory.

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Abnormal plasma enzyme activities

 Alanine Transaminase (ALT)/

glutamate pyruvate transaminase (GPT)
 Alanine aminotransferase (ALT)
 Very high values are seen in acute hepatitis, either
toxic or viral in origin.
 Both ALT and AST are increased in liver diseases, but
ALT >AST.
 Moderate increase may be seen in chronic liver
disease such as cirrhosis, and malignancy in liver.
 (AST/ALT) in normal conditions is 1.33 ± 0,42.
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Abnormal plasma enzyme activities

 Creatine Kinase (CK, CPK)

 Creatine Kinase (CK)
 High concentrations of CK in:
– skeletal muscle
– cardiac muscle
– brain tissue
 Increased plasma CK activity is associated
with damage to these tissues

  CK is especially useful to diagnose:

 AMI
 Skeletal muscle diseases ( Muscular Dystrophy )
Creatine Kinase isoenzymes
 CK occurs in 3 isoenzymes, each is a dimer
composed of 2 subunits (B & M): CK1 = BB, CK2 =
MB and CK3 = MM
 Normal serum consists of:
– Approximately 94% to 100% CK-MM
– Values for the MB isoenzyme range from undetectable to
trace (<6% of total CK).
– CK-BB is also present in small quantities
 Cardiac muscle CK is 80% CK-MM and 20% CK-MB
Creatine Kinase isoenzymes

 Each CK isozyme shows a characteristic

electrophoretic mobility.
Diagnostic Significance
 The value of CK isoenzyme separation can be
used principally in detection of myocardial
damage.
– increased CK – MB ( > 6% of the total CK
activity ) is a strong indication of AMI
 Post AMI
– CK-MB increases 4 – 8 hours
– Peaks at 12 - 24 hours
– Returns to normal 48 - 72 hours
Cardiac Disorders
 The CK rise the earliest, the LDH rise is latest
 The LDH elevatithose of CK and AST ons are
present longer than
Fold elevation

Days post infarction

Abnormal plasma enzyme activities
 α-Amylase

– hydrolyses alpha-bonds of large alpha-linked

polysaccharides such as starch and glycogen,
yielding glucose and maltose

– It is used as a marker to detect acute

pancreatitis and appendicitis
Abnormal plasma enzyme activities

 Gamma-glutamyl-transferase (GGT)

carboxypeptidase which cleaves C-terminal

glutamyl groups and transfers them to
peptides and other suitable acceptors
Abnormal plasma enzyme activities

 Alkaline Phosphatase (ALP)

– Widely distributed throughout the body
– High levels are seen is liver, bone, placenta and
intestine
– Physiological increases are been in pregnancy,
due to the placental isoenzyme, and in
childhood (when bones are growing), due to the
bone isoenzyme.
 Diagnostic Significance
 Alkaline Phosphatase (ALP)
– In hepatobiliary obstruction, hepatocytes lining
the biliary ducts induces the ALP synthesis.
– High levels of ALP is indicative of extrahepatic
obstruction rather than intrahepatic obstruction
– In bones, the enzyme is derived from osteoblasts.
Hence increased in bone diseases like rickets,
osteomalacia, neoplastic diseases with bone
metastates and healing fractures
Abnormal plasma enzyme activities

 Acid Phosphatase (ACP)

– ACP is secreted by prostate cells, RBC, platelets and
WBC.
– The main source of ACP is prostate gland and so can
be used as a marker for prostate disease.
– Different forms of acid phosphatase are found in
different organs, and their serum levels are used as a
diagnostic for disease in the corresponding organs.
Abnormal plasma enzyme activities
L a c ta te D e h y d ro g e n a s e
 LDH O O  O O 

C NADH + H+ NAD+ C
C O HC OH
CH 3 CH 3
p y ru v a te la c ta te

– High activities in heart, liver, muscle, kidney, and

RBC
– Lesser amounts: Lung, smooth muscle and brain
Abnormal plasma enzyme activities
 LDH
– LDH occurs in 5 isoenzymes: LDH1 (H4),
LDH2 (H3M), LDH3 (H2M2), LDH4 (HM3)
and LDH5 (M4)
Abnormal plasma enzyme activities
 LDH
– LDH occurs in 5 isoenzymes:
 LDH1 (H4): Cardiac , RBCs
 LDH2 (H3M): Cardiac , RBCs
 LDH3 (H2M2): Lung, spleen, pancreas
 LDH4 (HM3): Hepatic
 LDH5 (M4): Skeletal muscle
Diagnostic Significance
 LDH is elevated in a variety of disorders:
– in cardiac,
– hepatic,
– skeletal muscle,
– and renal diseases,
– as well as in several hematologic and neoplastic
disorders
 The highest levels of LD-1 are seen in pernicious
anemia and hemolytic disorders
 LD-3 with pulmonary involvement
 LD-5 predominates with liver & muscle damage
 Diagnostic Significance

 In healthy individuals
– LD-2 is in highest quantity then LD-1, LD-3, LD-4
and LD-5
 Heart problems:
– If problem is not MI, both LD1 and LD2 rise,
with LD2 being greater than LD1
– If problem is MI, LD1 is greater than LD2.
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 Diagnostic Significance

 LDH-6 has been present in patients with

arteriosclerotic cardiovascular failure
 Its appearance signifies a grave prognosis
and impending death
 It is suggested, that LDH-6 may reflect liver
injury secondary to severe circulatory
insufficiency
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Abnormal plasma enzyme activities

 Lipase
It is highly elevated in acute pancreatitis
and this persists for 7-14 days. Thus, lipase
remains elevated longer than amylase.
 Enzymes as Tumor Markers
Enzyme Disease
Serum acid phosphatase Prostrate cancer
Serum Alkaline phosphatase  Metastasis in liver, jaundice due
to carcinoma head of pancreas,
osteoblastic metastasis in bones
Serum LDH Advanced malignancies and
Leukemias
β- Glucuronidase Cancer of urinary bladder
Leucine Amino Peptidase (LAP) Liver cell carcinoma
Neuron specific Enolase  Malignancies of nervous tissue
and brain 
 Major Enzymes of Clinical
Significance
Disease Enzyme
Cardiac Disorders AST-LDH1-CK
Hepatocellular Disorders
Viral hepatitis: Hepatitis B & Hepatitis C.
Toxic hepatitis: caused by chemicals &
ALT-AST-LDH5
Toxins
Skeletal Muscle Disorders CK-AST
Biliary tract disorders ALP- GGT
Bone Disorders ALP
Acute Pancreatitis Lipase-AMS
Salivary Gland Inflammation AMS
References

 Victor A Hoffbrand, Paul Moss, J Pettit; Essential

Haematology. Essentials Series Blackwell Science,
New York; 2008.
 Victor W. Rodwell, David A. Bender, Kathleen M.
Botham, Peter J. Kennelly, P. Anthony Weil. Harper’s
Illustrated Biochemistry. McGraw-Hill Ed, 31 ed,
2018.