Cell Signaling/ Biosignalling

 Biochemistry & Genetics-II

 Course Code: SHS.109
 Pre-requisite: 103
 Credit Hours: 3
Lecture by

Dr. Naveed Munir

PhD Biochemistry (GCUF & KCL, UK)
M.Phil, M.Sc Biochemistry (UAF)
B. Sc Med. Lab Technology (UHS)
Member Pakistan Association of Medical Lab scientists (MPAMLS)
REFERENCE TEXT BOOK:
Lippincott’s Illustrated Review of Biochemistry by Pamela C. Champe and Richard A. Harvey Latest Ed.
Lehninger Principles of Biochemistry Fourth Edition, 2001. David L. Nelson (U. of Wisconsin–Madison) Michael M.
Cox (U. of Wisconsin–Madison)
 Urea cycle &
Specialized products made
from amino acids
 Conti.

 Ammonia metabolism

 Urea cycle

 Reactions involved in urea cycle

 Importance of urea cycle

 Fate of urea

 Specialized products made from Amino Acids

Blood level: Normally < 0.1 mg / dl

Urine level: 0.7 gm / day

Sources & fate (Ammonia):

 Various nitrogenous compounds
Amino acids
as

Purines& pyrimidines Some

(amino groups attached neurotransmitters Urea secreted into
Deamination: to the rings) as the intestine
--Oxidative deamination
--Transdeamination Monoamines:
--Specific deamination --serotonin
Methods
Histamine
Catabolism –epinephrine,norepinephrine,
( MAIN SOURCE) dopamine & their metabolites
metanephrine, normetanephrine Histaminase
&3 methoxy tyramine Intestinal bacterial
urease
Monoamine oxidase
(MAO)

NH3
Synthetic Catabolic
pathway pathway
Liver

Transdeamination (MAIN FATE)

Non essential glutamine Excreted

Urea
a.a.synthesis in urine
Urine
Transport of Ammonia to Liver
 Ammonia has to be eliminated
 Ammonia originates in the catabolism of amino acids
that are primarily produced by the degradation of
proteins – dietary as well as existing within the cell:
 Digestive enzymes
 Proteins released by digestion of cells sloughed-off
from the walls of the GIT
 Muscle proteins
 Hemoglobin
 Intracellular proteins (damaged, unnecessary)
 Ammonia has to be eliminated

• Ammonia is toxic, especially for the CNS, because it

reacts with -ketoglutarate, thus making it limiting
for the TCA cycle  decrease in the ATP level.

• Liver damage or metabolic disorders associated with

elevated ammonia can lead to tremor, slurred
speech, blurred vision, coma, and death.
 Ammonia has to be eliminated

 The two major types of hyperammonemia are:

1. Acquired hyperammonemia
 Liver disease is a common cause of hyperammonemia in
adults, and may be due, for example,
 To viral hepatitis or to hepatotoxins such as alcohol.

 Cirrhosis of the liver

2. Congenital hyperammonemia
 Genetic deficiencies of each of the five enzymes of the urea
cycle have been described in next section, with an overall
prevalence estimated to be 1:25,000 live births.
Also known as “Ornithine cycle”

Krebs’ Henseleit Urea Cycle – firstly described in 1932

Urea
 UREA CYCLE

 Urea is the major disposal form of amino groups derived

from amino acids, and accounts for about 80 - 90% of the
nitrogen-containing components of urine.

 Cycle of biochemical reactions occurring in many animals

 In mammals, the urea cycle takes place primarily in the

liver, and to a lesser extent in the kidney.
 UREA CYCLE

 Urea is synthesized in the liver

 Than secreted into blood stream

 And taken up by the kidneys for excretion in the urine

 Synthesis partly takes place in mitochondria and partly in

cytoplasm.
 UREA CYCLE

 The overall reaction of the urea cycle is as follow

NH4+ + HCO3- + H2O + 3ATP + Aspartate Urea + 2ADP + AMP + 2Pi

+ PPi + Fumarate
 STEPS OF UREA CYCLE

1. Formation of carbamoyl phosphate

Enzyme Carbamoyl Phosphate synthetase-I
Take place in mitochondrial matrix
 STEPS OF UREA CYCLE

2. Formation of citrulline
 Enzyme Ornithine transcarbamoylase
 Take place in mitochondrial matrix
 Citruline transported to cytosole
 STEPS OF UREA CYCLE

3. Formation of argininosuccinate
Enzyme Argininosuccinate synthetase
Take place in cytoplasm
Aspartate provide second NH3 group
 STEPS OF UREA CYCLE

4. Formation of Arginine & fumarate

Enzyme Argininosuccinase
Take place in cytoplasm
Fumarate utilize in citric acid cycle (Linkage)
 STEPS OF UREA CYCLE
5. Formation of Urea from Arginine
Enzyme Arginase
Take place in cytoplasm
Ornithine is also synthesized

Arginase
+ H2O
+
o
Urea
 Steps
Mitochondria
CO2 + NH3
2ATP
Carbamoyl-phosphate synthetase 1
1 2 ADP+1 Pi
O O
ll ll
H2N – C – O – P – OH Carbamoyl-phosphate
l CH2–CH2–CH2–CH–COOH
OH 2 Pi
l l
CH2–CH2–CH2–CH–COOH Ornithine NH NH2
l l transcarbamoylase l
Ornithine Citrulline
NH2 NH2 O=C–NH2

HOOC–CH2–CH–COOH
O Ornithine Citrulline l
ll NH2
Aspartic acid
H2N–C–NH2 Urea
Arginase Arginino-succinate 3 ATP
synthetase
5
CH2–CH2–CH2–CH–COOH AMP+PPi
l l H2O H2O
CH2–CH2–CH2–CH–COOH
NH NH2
l l
l Arginine Arginino succinate
NH NH2
NH2–C=NH
l
Arginino succinase NH–C=NH
l
4 Cytoplasm
HOOC–C–H Fumarate HOOC–CH2–CH–COOH
ll
H–C–COOH
 STEPS OF UREA CYCLE

1- C & O: from CO2

O
2- 1st N atom: from NH3
ll
3- 2nd N atom: from aspartate H2N–C–NH2

There in no net gain or loss of ornithine,

citrulline , argininosuccinate or arginine.

Ornithine is regenerated with each turn of

the urea cycle..
 Excretion of Urea

 Urea formed in liver goes to blood stream and excreted by

Kidneys

 A portion of the urea diffuses from the blood into the

intestine, and is cleaved to CO2 and NH3 by bacterial
urease. This ammonia is partly lost in the feces, and is
partly reabsorbed into the blood.

 Disorders of Liver or Kidney disturb the blood urea level.

 Normal range 20 to 40 mg/dL

CATABOLISM OF THE CARBON
SKELETONS OF AMINO ACIDS
GLUCOGENIC AND KETOGENIC AMINO
ACIDS
 GLUCOGENIC AND KETOGENIC AMINO
ACIDS

A. Glucogenic amino acids

 Amino acids whose catabolism yields pyruvate or
one of the intermediates of the citric acid cycle are
termed glucogenic.
 These intermediates are substrates for
gluconeogenesis and, therefore, can give rise to the
net formation of glucose in the liver and kidney.
 GLUCOGENIC AND KETOGENIC AMINO
ACIDS
B. Ketogenic amino acids
 Amino acids whose catabolism yields either acetoacetate
or one of its precursors (acetyl CoA or acetoacetyl CoA)
are termed ketogenic.

 Acetoacetate is one of the ketone bodies, which also

include 3-hydroxybutyrate and acetone.

 Leucine and lysine are the only exclusively ketogenic

amino acids found in proteins.
Catabolism of the Carbon Skeletons
of Amino Acids
 Anabolism

Conversion of Amino Acids

to Specialized Products
 Importance

 In addition to serving as building blocks for proteins, amino

acids are precursors of many nitrogen-containing

compounds that have important physiologic functions
 Porphyrins

 Porphyrins are cyclic compounds that readily bind

metal ions—usually Fe2+ or Fe3+.

 Heme

Prosthetic group of:

 Hemoglobin

 Myoglobin

 Cytochromes

 Catalase

 Nitric oxide synthase

 Peroxidase
 Porphyrias

 These hemeproteins are rapidly synthesized and degraded.

 Porphyrias are rare, inherited (occasionally acquired)

defects in heme synthesis,

 Resulting in the accumulation and increased excretion of

porphyrins or porphyrin precursors

 “Porphyria” refers to the reddish/ purple color caused by

pigment-like porphyrins in the urine of some patients with

defects in heme synthesis.
 Clinical manifestation

 Photosensitivity—that is, the skin itches and burns

(pruritus) when exposed to visible light

 Reactive oxygen species can oxidatively damage

membranes, and cause the release of destructive enzymes
from lysosomes
 Skin eruptions in a patient with
Porphyria Cutanea tarda (PCT)

Painful, blistering skin lesions that develop on sun-exposed skin (photosensitivity)

 Neurotransmitters (Catecholamine)
 Catecholamines

Catecholamines help the body respond to stress or fright

and prepare the body for "fight-or-flight" reactions.
 Dopamine,

 Norepinephrine, and

 Epinephrine

are biologically active (biogenic) amines that are collectively

termed catecholamines.
 Neurotransmitters (Catecholamine)

 Dopamine and norepinephrine are synthesized in the

brain and function as neurotransmitters

 Norepinephrine is also synthesized in the adrenal

medulla, as is epinephrine.
Synthesis of catecholamine
 Parkinson disease

 A neurodegenerative movement disorder, is due to

insufficient dopamine production as a result of the
idiopathic loss of dopamine-producing cells in the brain.
 Histamine

 Histamine is a chemical messenger and a powerful vasodilator, that

mediates a wide range of cellular responses,

 Including allergic and inflammatory reactions,

 Gastric acid secretion, and

 Possibly neurotransmission in parts of the brain.

 Agents that interfere with the action of histamine have important

therapeutic applications
 Secreted by white blood cells (mast cells) as a result of allergic reactions

or trauma
Biosynthesis of Histamine
 HISTIDINEMIA
 The disease is due to a deficiency in histidase.

 Elevated levels of histidine occur in blood and urine.

 Defect in speech and mental retardation

 Serotonin
 Serotonin, also called 5-hydroxytryptamine (5HT), is
synthesized and stored at several sites in the body
including GI tract (90%), platelets (8%), brain (1-2%) and
other parts .
 By far the largest amount of serotonin is found in cells of
the intestinal mucosa in the enteric nervous system located
in the gastrointestinal tract (GI tract) to regulate intestinal
movements.

 Smaller amounts occur in the central nervous system,

where it functions as a neurotransmitter, and in platelets.
 Functions
Key hormone that stabilizes our mood, feelings of well-being,
and happiness.
 Pain perception
 Regulation of sleep
 Appetite
 Temperature
 Blood pressure
 Cognitive functions
 Including learning, thinking, reasoning, remembering, problem
solving, decision making, and attention
Serotonin Synthesis
 Melanin

 Melanin is a pigment that occurs in several tissues,

particularly the eye, hair, and skin.

 It is synthesized from tyrosine in the epidermis by pigment-

forming cells called melanocytes

Melanin
 Functions

 Its function is to protect underlying cells from the harmful

effects of sunlight.

 A defect in melanin production results in albinism, the most

common form being due to defects in copper-containing

tyrosinase also known as tyrosine hydrolase.
 Purines and Pyrimidine
Nitrogenous Bases
 One of the important specialized pathways of a number of
amino acids is the synthesis of purine and pyrimidine
nucleotides.
 Purines consist two ring structure includes Adenine and
Guanine
 Pyrimidines single ring includes Uracil, Thymine,
Cytosine
 Source of energy (ATP, GTP), Part of Coenzymes (NAD),
synthesis of DNA & RNA etc
Purines and Pyrimidine
Nitrogenous Bases
 HOME TASK!

Make ten mcq’s from lecture....

How can you differentiate transamination and

deamination.

Enlist all enzymes involved in protein digestion

with their functions.