Dr. Vijetha Shenoy Belle Associate Professor & In-Charge Clinical Biochemistry Laboratory Department of Biochemistry KMC Manipal

DR.
VIJETHA SHENOY BELLE
Associate Professor & In-charge Clinical Biochemistry Laboratory

Department Of Biochemistry
KMC Manipal
OVERVIEW
 imparts competency based education
 conduct good quality research- ICMR projects
 provide high standard of diagnostic services

RECOMMENDED BOOKS
Textbook of Biochemistry, D.M Vasudevan,

Sreekumari & K. Vaidyanathan
Biochemistry by U.Satyanarayana
Modes of teaching
 Lectures (Theory classes)
 Small group teaching (SGT)
 Self directed learning (SDL)
 DOAP (Practical)
 Early clinical exposure
• A 100 page long book for writing SGT, SDL & ECE
• Record book for practical
BIOCHEMISTRY
 Carl A Neuberg
 1903
BIOCHEMISTRY
“BIOS” = Life
The chemistry of “LIFE”
The science concerned with the

chemical basis of life
BIOCHEMISTRY
Our body is a huge LABORATORY

LIFE
What we call LIFE is due to
the biochemical reaction taking
place in the body
Why have so many reactions?
To give ENERGY
To GROW
To REPRODUCE
To heal and repair worn out
Tissues
To keep the body healthy
To fight against disease causing

organisms
Basic elements in the cell
Carbon
Hydrogen
Oxygen
Nitrogen
Phosphorus
Sulphur,
Calcium,
Magnesium
Livi
ng
Basic molecules in the cell
Carbohydrates
Lipids
Proteins
Nucleic Acids
Water
Living or
non-living???
CEL
L
B S of 1 0 m o n t h old
B o y
Living or non-living?????
BIOCHEMISTRY
Human Biochemistry
Plant Biochemistry
Microbial Biochemistry
Biotechnology
Genomics
Proteomics
Bioinformatics
Our focus in MBBS
HUMAN BIOCHEMISTRY and NUTRITION

• 1944—DNA is shown to be the
hereditary material
• 1953—Concept of the double helix is
posited
• 1966—The genetic code is solved
• 1972—Recombinant DNA
technology
is developed
• 2003—Sequencing of the first
human
Clinical Biochemistry
Applies basic biochemistry and analytical chemistry to

medical diagnosis, treatment and management.
Biochemical diagnosis
Heart Disease
Liver Disease
Detection of cancer
Electrolyte imbalance
Blood Gas Analysis
Hemoglobin abnormalities
Molecular biology diagnostics
Metaphase
Karyotype
Learning objective
Correlate biochemical pathogenesis to
- Clinical Manifestations
- Laboratory investigation results
- Natural history of the disease
- Complications
Learning objective (Competency BI 1.1)
• Understand
the importance of various macromolecular
components of the cell and their functions
• Understandthe structure, function and inter-relationships

of biomolecules and consequences of deviation from
normal (biochemical aspects with few examples in
general)
Cell
• structural and functional unit of life
• basic unit of biological activity

Cell and subcellular organelles
 Separated by differential ultracentrifugation
 Based on density fractions form pellets and the

supernatant is formed
This supernatant has cytoplasm –

The marker enzyme is Lactate Dehydrogenase
Subcellular Pellet formed at centrifugal Marker enzyme
organelle force
Nucleus 600-750 x g (10 min)
Mitochondria 10000-15000 x g (10 min) Inner membrane: ATP synthase
Lysosome 18000-25000 x g (10 min) Cathepsin
Golgi complex 35000-40000 x g (30 min) Galactosyl

transferase
Microsomes 75000-100000 x g (100 min) Glucose 6-

phosphatase
Cytoplasm Supernatant Lactate

dehydrogenase
HOMEWORK ….
 Functions of cell organelles
 Differences between cytoplasm and cytosol

Lysosomes
tiny organelles; meant for degradation of : proteins, carbohydrates, lipids
and nucleotides
Lysosomal enzymes: hydrolyzing enzymes, optimum pH
around 5.
Polysaccharides hydrolyzing enzymes: Glucosidase, galactosidase,

mannosidase, hyaluronidase, sulfatase
Protein hydrolyzing enzymes- cathepsins, collagenase, elastase, peptidase
Lipid hydrolyzing enzymes - phospholipases
When membrane is disrupted, the released enzymes can hydrolyze external

substrates leading to tissue damage
1. Lysosomal storage diseases
Lysosomal enzymes are deficient leading to accumulation of lipids or

polysaccharides
2. Inclusion cell (I-cell) disease
Rare, lysosomes lack enzymes, but they are seen in blood
Mannose-6-phosphate is absent –This is a marker to target the nascent enzymes to

lysosomes
Enzymes can not reach their destinations-non entry of enzymes into lysosomes
Protein targeting defect

3. Cathepsins – Role in tumor metastasis
Cancer cells liberate the cathepsins out of the cells, which
degrade the basal lamina by hydrolyzing collagen and elastin,
so that other tumor cell can travel out to form metastasis
Marker enzyme - Cathepsins

4. Gout : when the urate crystals are
phagocytosed, cause physical damage to lysosomes, and release
of enzymes inflammation and arthritis result
Peroxisome
They contain peroxides and catalase, help in destroying

unwanted peroxides and other free radicals
• Insufficient oxidation of very long chain fatty acids by
peroxisomes- adrenoleukodystrophy (deficiency of peroxisomal
matrix proteins)
• Characterized by progressive degeneration of liver, kidney, brain
• Autosomal recessive condition

• In Zelweger syndrome, proteins are not transported to
peroxisomes
ECM – extra cellular
matrix
SLO
 At the end of this lecture class student should know

1. What is extracellular matrix?
2. List the components of ECM
3. Functions
SLO 1- What is extra cellular matrix?
 Cell and organs are surrounded by a glue like substance called ECM
 Constitutes the non-cellular components of all tissues and organs
 Mainly – supporting structure for the framework of the tissues
 Actually plays – part – development, differentiation and maintenance of the tissues

and organs
SLO 2 - List the components of ECM
 Water
 Proteins
 Polysaccharides
 Proteoglycans
 Fibrous proteins
Components of ECM are unique – function
ECM – highly dynamic structure – constantly being remodeled

SLO 3: Functions of ECM
 Functional properties of ECM depends on the molecular compositions
Main
components
Proteoglycans Fibrous protein
 The ECM is composed of an interlocking mesh of fibrous proteins and GAGs

 Biochemical
and mechanical properties of each organ – dependent on
ECM composition
 Post translational modifications undergone by ECM proteins are also

significant in their functions
Proteoglycans of ECM
 Major protein
 GAG chains linked to specific protein core
1. Leucine-rich proteoglycans – small molecules

2. Cell surface glycoproteins – act as core receptors – signal transduction
pathways involving growth factors + receptors
3. Modular glycoproteins – mediate cell adhesion migration and proliferation
The major proteins of ECM
 Collagen
 Elastin
 Fibrillin
 Fibronectin
 Laminin
Collagen
 To give support to organs

 To provide alignment of cells – cell anchoring is possible
 Proliferation and differentiation of cells
 In blood vessels – collagen is exposed – platelet adhere –
thrombus formation
Elastin
 ECF protein – recoil to tissues that undergo repeated stretching –

Elasticity
 Major component of elastic fibers
 Found in ligaments and Walls of the blood vessels

Fibrillin
 Is a large glycoprotein which is essential for the formation of elastic fibres found
in connective tissue.
 Is a component of myofibril
 Found in eye lens, periostium of the bone and elastin fibres of aorta
 Deficiency: Marfan syndrome

Fibronectin
 A glycoprotein
 Cell adhesion and migration
 Clinical importance: Tumor cells that are deficient in fibronectin –

lack of adhesion – lead to metastasis
Laminin
 Basal lamina of glomerular membrane
 First Extracellular protein synthesized during embryogenesis
 Involved in neuronal growth and nerve regeneration
 Alzheimer’s disease
SLO covered???
 Define ECM?
 Components?
 Functions?
Questions for SGT
 Explain the structure of cell membrane

 Explain the transport across the cell membrane
 What is extracellular matrix? List the components of ECM
 Functions of ECM
THANK YOU

Dr. Vijetha Shenoy Belle Associate Professor & In-Charge Clinical Biochemistry Laboratory Department of Biochemistry KMC Manipal

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DR.

VIJETHA SHENOY BELLE

Associate Professor & In-charge Clinical Biochemistry Laboratory

 imparts competency based education

 conduct good quality research- ICMR projects

 provide high standard of diagnostic services

Textbook of Biochemistry, D.M Vasudevan,

The chemistry of “LIFE”

The science concerned with the

Our body is a huge LABORATORY

To heal and repair worn out

To keep the body healthy

To fight against disease causing

HUMAN BIOCHEMISTRY and NUTRITION

Applies basic biochemistry and analytical chemistry to

Correlate biochemical pathogenesis to

- Laboratory investigation results

- Natural history of the disease

• Understandthe structure, function and inter-relationships

• structural and functional unit of life

• basic unit of biological activity

 Based on density fractions form pellets and the

This supernatant has cytoplasm –

Nucleus 600-750 x g (10 min)

Mitochondria 10000-15000 x g (10 min) Inner membrane: ATP synthase

Lysosome 18000-25000 x g (10 min) Cathepsin

Golgi complex 35000-40000 x g (30 min) Galactosyl

Microsomes 75000-100000 x g (100 min) Glucose 6-

Cytoplasm Supernatant Lactate

 Functions of cell organelles

 Differences between cytoplasm and cytosol

Polysaccharides hydrolyzing enzymes: Glucosidase, galactosidase,

Protein hydrolyzing enzymes- cathepsins, collagenase, elastase, peptidase

Lipid hydrolyzing enzymes - phospholipases

When membrane is disrupted, the released enzymes can hydrolyze external

Lysosomal enzymes are deficient leading to accumulation of lipids or

Mannose-6-phosphate is absent –This is a marker to target the nascent enzymes to

Protein targeting defect

Marker enzyme - Cathepsins

They contain peroxides and catalase, help in destroying

• Characterized by progressive degeneration of liver, kidney, brain

• Autosomal recessive condition

 At the end of this lecture class student should know

 Constitutes the non-cellular components of all tissues and organs

 Mainly – supporting structure for the framework of the tissues

 Actually plays – part – development, differentiation and maintenance of the tissues

Components of ECM are unique – function

ECM – highly dynamic structure – constantly being remodeled

Proteoglycans Fibrous protein

 The ECM is composed of an interlocking mesh of fibrous proteins and GAGs

 Post translational modifications undergone by ECM proteins are also

1. Leucine-rich proteoglycans – small molecules

 To give support to organs

 ECF protein – recoil to tissues that undergo repeated stretching –

 Major component of elastic fibers

 Found in ligaments and Walls of the blood vessels

 Deficiency: Marfan syndrome

 Cell adhesion and migration