Metabolism of Proteins

1. Nitrogen Balance
2. Digestion & Absorption of
Proteins

Mbbs/BDS/March 2019

1
Nitrogen Balance

2
 Overview

• Amino acids are not stored

• Used for protein synthesis
• Sources – dietary proteins
– denovo synthesis
– degradation of proteins
• Excess amino acids are degraded & reutilized

3
 Catabolism overview

Carbon skeleton acts

Removed as NH3
as intermediates of
Transamination &
energy producing
Oxidative deamination
metabolic pathways

4
 Amino Acid Pool

• 100 g in an avg adult

• Circulating free amino acids
• Source – diet, protein breakdown, denovo synthesis
• Fates – protein synthesis, TCA, specialized products

5
Nitrogen Balance

6
 Protein Turnover

• Breakdown and synthesis of proteins

• In healthy individuals synthesis equals breakdown
• Variable for different proteins
• Depends on half life

7
 Rate of protein turnover
• Protein turnover leads to daily hydrolysis and resynthesis of about 200
g of body proteins.

• An average adult degrades 1-2% of the total proteins everyday, the

major proportion of which is the muscle proteins.

• starvation also stimulates protein degradation.

• protein turnover varies according to the physiological state, i.e., the

half-life of proteins in specific tissues responds to the physiological
requirement of the organism.

• The proteins in tissues undergoing development / differentiation /

major structural rearrangement (e.g., uterus during pregnancy) have
shorter half-lives.

8
 Half life

• The half-lives of proteins vary from 30 seconds to many days to yrs

• Enzymes have a high turnover
• Hemoglobin has a very long half-life and mostly lasts for the life of
the RBC (about 110 days).
• Structural proteins such as collagen are metabolically stable and have
half-lives in months or years.
• Half-life of mature functional proteins is related to the N-terminal
AA, where, some AAs when present on the N-terminal stabilize the
proteins and hence increase their half-lives, e.g., Met, Ser, Ala, Thr,
Val and Gly.
• Certain AAs on the N-terminal might decrease the half-life of
proteins, e.g., Phe, Tyr, Leu, Ile, Lys, Arg, Asp, and Glu.

9
 Signals for protein degradation
• Half life of proteins determined by N-terminal amino
acid sequence

• Proteins with sequences Proline ,Glutamate , Serine

,Threonine (PEST) – Short lived
 Protein
degradation

Ubiquitin-
Lysosomal
Proteasome
enzymes
complex
• Degrades intracellular proteins.
• Degrades circulating plasma
11
proteins.
 Ubiquitin-Proteasome degradation

• Ubiquitin is a small globular protein

• Proteins destined to be degraded are linked with ubi.
• Ubiquitin is linked to the protein through ϵ-amino gp of a
lysine moiety
• Labelled proteins are recognized by a large barrel shaped
proteolytic molecule called as proteasome

12
Ubiquitin (76 AA )

This ‘ubiquitination’ of proteins is dependent upon the N-

terminal AA, i.e., N-End rule.
Met and ser prevent ubiquitination, whereas, asp and arg
accelerate the process.

Ubiquitinated proteins are delivered to be degraded by a large

complex known as the 26S proteasome. The proteasome
consists of two proteins; regulatory and catalytic - each of at
least 32 different subunits.

13
Ubiquitin – Proteasome complex

14
 Nitrogen balance
Primarily, proteins serve as the building blocks of the body
and only about 15% of the energy is derived from proteins.
Amino acids may serve as the exclusive sources of energy
during starvation.
The minimum level of intake of proteins is 0.75 g/kg body
weight per day.

• A normal healthy individual is said to be in nitrogen

balance state, as the intake roughly equals the output via
urine, skin and feces.

15
• Negative Nitrogen Balance: The balance is said to be
negative, if the output exceeds the intake.

• Positive Nitrogen Balance: The balance is said to be

positive, if the intake exceeds output.

16
 Factors affecting N balance

1. Dietary Protein deficiency. The deficiency of one or

more amino acids may cause negative nitrogen balance.

17
 Factors affecting N balance

• 2. Physiological state.
• Starvation is a major cause of disturbance in nitrogen
balance. Similarly, the balance is negative during
malignancy, uncontrolled diabetes mellitus and other chronic
diseases.
• Convalescence after an illness or surgery is accompanied by
tissue regeneration and, hence, a positive nitrogen balance.
• During the active growth period, nitrogen balance is positive
due to tissue building up.
• A pregnant woman is also under state of positive nitrogen
balance due to protein retention for the growth of fetus.
18
 Factors affecting N balance

• 3. Hormonal status. Different hormones have different

impact on nitrogen balance,
• e.g., growth hormone, insulin and androgens promote
positive nitrogen balance,
• corticosteroids promote negative nitrogen balance.

19
Digestion & Absorption of
Proteins

20
 Digestion

• Must be hydrolyzed first to free AAs, which can be

absorbed. About 95-99% of ingested proteins are
digested and absorbed in GIT.
• No protein digestion in the mouth as saliva lacks any
protease.

21
Digestion of protein - hydrolysis
NH2 H
R CH C N CH COOH

O +H2O R

+ proteases
NH2
HHN CH COOH
R CH C OH
R
O
23
 Hormones regulating protein digestion

• Gastrin: Secreted from gastric antrum and duodenum

mucosal cells in response to the entry of food bolus into
the stomach and stimulates secretion of pepsinogen and
the intrinsic factor from gastric mucosa.
• Cholecystokinin (CCK): Released from duodenum and
jejunum and activates pancreatic proenzyme secretion.
• Secretin: Located in duodenum and jejunum and
stimulates secretion of bicarbonate-rich pancreatic juice.

24
 Protein digestion begins in stomach

• Gastric pH denatures proteins

• HCL – parietal cells, makes proteins more
susceptible to proteolytic cleavage

• Pepsinogen is activated to pepsin by gastric acidity

25
Formation of HCl in parietal cells

26
• Pepsin - inactive precursor pepsinogen
• Active @ pH 2-3, inactive pH>5
• Secretion stimulated by acetylcholine or
acid
• Only protease which can break down
collagen
• Action terminated by neutralisation by
bicarbonate in duodenum.
 Digestion by pancreatic enzymes

• Products of pepsin degradation are cleaved by panc enz

• 4 enzymes – elastase, carboxypeptidase, trypsin and
chymotrypsin
• All secreted as zymogens – activated and released by 2
GI hormones – Secretin & Cholecystokinin
• Enteropeptidase activates Trypsin by cleaving a 6 aa
long peptide at amino terminus of trypsinogen
• Trypsin then activates other enzymes

28
Activation of pancreatic proteases
Enterokinase

Trypsinogen Trypsin

Trypsinogen Trypsin
Chymotrypsinogen Chymotrypsin
Proelastase Elastase
Procarboxypeptidase Carboxypeptidase
 Specificity of Pancreatic enzymes

• Each of these enzymes have a specific location for its

cleavage of peptide bond

30
 Enzymes of the intestine

• Aminopeptidase – secreted by intestinal epithelium

• Cleaves N-terminal amino acid of oligopeptides
• Free amino acid is released and absorbed

31
 Absorption

• Free amino acids and dipeptides are absorbed in the

intestinal mucosal cell
• Dipeptides cleaved into a. a. inside the cell
• Only free amino acids are then absorbed into the portal
circulation and taken to the liver

32
 Absorption of whole protein without degradation

• Few cases
a) Normally in infants where -globulins (antibodies)
present in colostrum are absorbed to acquire passive
immunity by the baby .

b) Abnormally in adults in certain diseases that lead to

allergic reaction e.g. urticaria.
• Site of Absorption: Most of the absorption of AAs takes
place in jejunum

33
 Transport of amino acids into the cells

• Transport systems are required

• Specific for a set of amino acids
• 7 transport systems known so far
• One transport system has great clinical importance –
COLA transport system – cystine/Orn/Lys/Arg
• Reabsorbs these amino acids in the kidneys
• Absence leads to loss of these amino acids in urine
• Cystinuria

34
 Cystinuria

• 1 in 7000 incidence
• Most common
disorder of amino
acid transport
• Leads to cystine
stones in the
urinary tract

35
 Hartnup disease
• An autosomal recessive inherited disease. .

• The defect lies in the transport of neutral amio acids in

intestine, kidneys and brain. This leads to defective
transport of neutral AAs, particularly trp.

36
 Clinical featurs – Hartnup disease

• Symptoms : mental retardation, intermittent cerebellar

ataxia , other neurological symptoms , Pellagra-like skin
rash and hypersensitivity to sunlight. Most of these
symptoms are caused by the reduced synthesis of
serotonin and nicotinic acids due to excessive loss of trp.
• Blood levels of trp and other neutral AAs are lower than
normal.
• Fecal and urine content is 5 - 10 times higher than
normal (neutral aciduria).

37