Escolar Documentos
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Dr.Thomas Nyambo
Department of Biochemistry
School of Medicine MUCHS
Hormone
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Mechanism of action
Hormones are normally present in the plasma and
interstitial tissue at concentrations in the range of 10-
7M to 10-10M.
.
The G protein cycle
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G proteins and Signal transduction
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cAMP
.
Metabolism of cAMP
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cAMP as a second messenger
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Activation of protein kinase A by cAMP
Signal amplification
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Signal transductional pathways of the
Upon binding of FSH to the FSH receptor, the Gs subunit
FSH
dissociates.
Together with GTP, this complex directly activates adenylyl cyclase,
thereby leading to cAMP synthesis.
PKA is activated by cAMP, which causes the dissociation of the
catalytic subunit (C) from the regulatory subunit (R).
The active catalytic site can activate proteins by phosphorylation.
Conversely, the production of cAMP leads to an intracellular rise of
Ca2+, presumably due to the gating of calcium channels.
In the nucleus the catalytic subunit of the PKA can phosphorylate
transcription factors which then bind to CREs preceding certain
genes.
Finally, mRNA synthesis of primary response genes of FSH action
starts.
Signal transductional pathways of the FSH
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An Overview of Calcium Regulation in Cells
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The Action of Nitric Oxide on Blood
Vessels
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G proteins
G proteins mediate signal transduction through G
protein-linked receptors.
When a messenger binds to its receptor, the Gs
protein is activated.
In the inactive state, the alpha, beta, and gamma
subunits are present as a complex, with GDP bound
to the alpha subunit.
(a) When a receptor is activated by binding of its
specific ligand on the outer surface of the plasma
membrane, the receptor-messenger complex
associates with the Gs protein, causing the
displacement of GDP by GTP and the dissociation of
the Gs(alpha)-GTP complex.
G proteins
(b) The GTP-Gs complex then binds tightly to a molecule of
membrane-bound adenyl cyclase, activating it for synthesis
of cAMP.
(c) Activation ends when the ligand leaves the receptor, the
GTP is hydrolyzed to GDP by the GTPase activity of the
Gs subunit, and the Gs dissociates from the adenylyl
cyclase.
(d) Adenylyl cyclase then reverts to the inactive form, the Gs
alpha reassociates with the Gs complex, and cAMP
molecules in the cytosol are hydrolyzed to AMP by the
enzyme phosphodiesterase.
G proteins and Signal transduction
The cascade ends with activation of the final target enzyme
.
Phosphoinositide Cascade
Not all G proteins signal through adenylate cyclase .
Certain G proteins, such as Gq, act by activating phospholipase C.
When a particular receptor such as the vasopressin receptor is
activated, it activates a G protein that then activates the membrane-
bound enzyme phospholipase C (PLC).
Phospholipase C cleaves PIP2, releasing inositol 1,4,5-trisphosphate
(IP3) and diacylglycerol (DAG).
These two messengers then cause the activation of other proteins
Different isoforms of PLC are activated by different receptors by a
variety of mechanisms.
Examples of processes regulated by the phosphoinositide cascade are:
1) histamine secretion by mast cells
2) aggregation of blood platelets and release of serotonin
3) insulin secretion by pancreatic beta cells
4) smooth muscle contraction
IP3 and DAG
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IP3 and DAG linked cascade
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Insulin
.
Insulin
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Insulin Receptor
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Insulin signal transduction pathways
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GLUT4 translocation
Insulin promotes glucose uptake by muscle and
adipose tissue via stimulation of glucose transporter
(GLUT) 4 from intracellular sites to the plasma
membrane.
Attenuated GLUT4 translocation and glucose uptake
by muscle and fat cells following insulin stimulation
represent a prime defect in insulin resistance
The PI3 kinase/Akt pathway has been demonstrated
to be upstream of GLUT4 translocation.
The insulin receptor is a tyrosine kinase
One can say that it functions as an enzyme that
transfers phosphate groups from ATP to tyrosine
residues on intracellular target proteins.
Binding of insulin to the subunits causes the beta
subunits to autophosphorylate,thus activating the
catalytic activity of the receptor.
The activated receptor then phosphorylates a
number of intracellular proteins, which in turn alters
their activity, thereby generating a biological
response.
Actions of insulin on cell level
and global metabolism level
.
Glucagon
.
Hypothalamo-pituitary hormones
.
Hypothalamic/pituitary axis
Releasing factors produced by the hypothalamus are
Thyrotropin releasing hormone (TRH) releases thyrotropin (TSH)
Corticotropin releasing factor (CRF) releases adrenocorticotropin
(ACTH)
LH/FSH releasing hormone (GnRH) releases luteinizing hormone
(LH) and follicle-stimulating hormone (FSH)
Growth hormone releasing factor (GHRF) releases growth hormone
Prolactin releasing factor (PRF) releases prolactin
Melanocyte-stimulating hormone releasing factor releases
melanocyte- stimulating hormone (MSH)
Inhibitory factors produced by the hypothalamus are
Dopamine inhibits prolactin release
Somatostatin inhibits growth hormone and TSH release
Melanocyte-stimulating hormone inhibitory factor (MIF) inhibits
MSH release
Growth Hormone
.
Synthesis of the catecholamines from tyrosine
The first step in the process requires tyrosine hydroxylase in the
precence of tetrahydrobiopterin as cofactor.
The hydroxylation reaction generates DOPA (3,4-
dihydrophenylalanine).
DOPA decarboxylase converts DOPA to dopamine, dopamine b-
hydroxylase converts dopamine to norepinephrine and
phenylethanolamine
N-methyltransferase converts norepinephrine to epinephrine.this
reaction uses SAM as a methyl donor generating S-
adenosylhomocysteine.
In the substantia nigra and some other regions of the brain, synthesis
proceeds only to dopamine.
In the adrenal medulla dopamine is converted to norepinephrine and
epinephrine.
Catabolism of the catecholemines occurs through the actions of
catecholamine-O-methyltransferase, (COMT) and monoamine
oxidase, (MAO). Both of these enzymes are widely distributed
throughout the body. However, COMT is not found in nerve endings
Synthesis of the catecholamines from tyrosine.
.
Steroid hormones
The steroid hormones are all derived from
cholesterol, with the exception of retinoic acid
They all contain the same cyclopentanophenanthrene
ring and atomic numbering system as cholesterol,
with the exception of vitamin D, .
The conversion of C27 cholesterol to the C18, C19,
and C21 involves the rate-limiting, irreversible
cleavage of a 6-carbon residue from cholesterol.
This cleavage produces a C21(pregnenolone) and an
isocaproaldehyde.
Steroid hormones
Steroid hormones can be roughly divided into 4 groups
according to their physiological behaviour:
i. Adrenal mineralocorticoids, which regulate the salt
balance and maintain blood pressure.
ii. Glucocorticoids, which regulate carbohydrate metabolism
and manage stress,
iii. Progestogens and estrogens, which are mainly produced
by the ovaries and regulate reproductive function and
secondary sex characteristics in the female.
iv. Androgens, which are mainly testicular in origin and are
essential for fertility and secondary sex characteristics in
the male
Steroidogenic hormones
Most of the steroidogenic enzymes belong to the cytochrome
P450 oxidation group, among which five enzymes are
involved in adrenal steroidogenesis:
i. P450scc (side chain cleavage)
ii. P450c11 (11--hydroxylase)
iii. P450c17 (17--hydroxylase)
iv. P450c21 (21-hydroxylase)
v. P450aldo (aldosterone synthase).
The first two of these are located in the mitochondria and the
last two in the endoplasmic reticulum. In addition,
P450aro mediates the aromatisation of androgens to
estrogens in the gonads
Steroid hormones
Steroids with 21 carbon atoms are known
systematically as pregnanes
Those containing 19 and 18 carbon atoms are
known as androstanes and estranes,
respectively.
As already mentioned,retinoic acid and
vitamin D are not derived from pregnenolone,
but from vitamin A and cholesterol
respectively.
Steroid hormones
Pregnenolone: is produced directly from cholesterol,
the precusor molecule for all C18, C19 and C21 steroids
.
Progesterone: is a progestin which is produced directly
from pregnenolone and secreted from the corpus luteum,
responsible for changes associatedwith luteral phase of
the menstral cycle
.
Reactions of the zona glomerulosa
The zona glomerulosa cells lack the P450c17 that converts
pregnenolone and progesterone to their C17 hydroxylated
analogs.
In these cells the pathways to the glucocorticoids
(deoxycortisol and cortisol) and the androgens
(dehydroepiandosterone [DHEA] and androstenedione) are
blocked.
Zona glomerulosa cells contain the enzyme responsible for
converting corticosterone to aldosterone, the most potent
mineralocorticoid.
This enzyme is P450c18 (or 18a-hydroxylase), also called
aldosterone synthase.
Thus, the zona glomerulosa is responsible for the conversion
of cholesterol to corticosterone and aldosterone.
Reactions of zona fasciculata and zona reticularis
The cells of the zona fasciculata and zona reticularis
lack aldosterone synthase (P450c18) that converts
corticosterone to aldosterone, and thus these tissues
produce only corticosterone.
These two zones contain P450c17 and thus produce
the major glucocorticoid, cortisol.
Zona fasciculata and zona reticularis cells also
contain the C17,20 lyase, whose activity is
responsible for producing the androgens,
dehydroepiandosterone (DHEA) and
androstenedione.
Steroid Hormone Biosynthesis
Regulation of Adrenal Steroid Synthesis
.
Synthesis of the major female sex hormones in the ovary
.
Adrenal genital syndrome
Deficiency of 21-hydroxylase inhibits the synthesis of
glucocorticoids and mineralocorticoids .
This leadsto overproduction of testosterone in the
adrenal glands and underproduction of cortisol.
Underproduction of cortisol effectively increases
adrenocorticotropic hormone (ACTH), which
stimulates the adrenals to grow and synthesize
steroids, exacerbating the testosterone
overproduction.
This leads to masculinization of females.
Synthetic steroids
Synthetic steroid hormones include the
following:
Diethylstilbestrol, a synthetic estrogen
previously used to promote growth of beef
cattle, until it was found to be potentially
carcinogenic at the levels found in meat from
treated cattle.
Oral contraceptives, such as norethynodrel
and mestranol.
Anabolic Steroids
Were first developed in the 1930's by the Germans in the
second world war.
Then in the 1950's many athletes began to find that anabolic
steriods were beneficial to their physical strength
characteristics.
Pure Testosterone was the available drug, resulting in severe
side-affects which soon became apparent after prolonged use.
There after that, other choices like Dianabol or
Methandrostenolone were developed, and a decade later
anabolic steriods were available on the market.
Latest products are so called designer steroids such as
tetrahydrogestrinone (THG), which have been chemically
engineered to deceive anti doping testers.
.
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Congenital adrenal hyperplasia (CAH)
CAH is al term used to describe a group of inherited
disorders in which a defect in cortisol biosynthesis is
present.
An enzymatic defect in 11--hydroxylase is the
second most common variant of CAH.
Individuals with 11--hydroxylase deficiency present
with features of androgen excess, including
masculinization of female newborns and precocious
puberty in male children.
Approximately two thirds of cases also have
hypertension, which may or may not be associated
with mineralocorticoid excess, hypokalemia, and
metabolic alkalosis.
17-hydroxylase deficiency
Hypothyroidism in Adults
Low serum thyroid hormone resulting from hypothalmic, pituitary, or
thyroid insufficiency. Untreated, can lead to life-threatening myxedema
coma (early signs of fatigue, forgetfulness, sensitivity to cold,
unexplained weight gain, constipation progress to decreasing mental
stability, dry, flaky inelastic skin; puffy face, hands, and feet; hoarseness;
periorbital edema; upper eyelid droop; dry, sparse hair; and thick, brittle
nails.
Hypothyroidism in Children (Cretinism)
Deficiency of thyroid hormone secretion during fetal development or
early infancy results in infantile cretinism (congenital hypothyridism).
Respiratory difficulties, persistent jaundice, and hoarse crying in infants;
stunted growth (dwarfism), bone and muscle dystrophy, and mental
deficiency in older children.
Thyroid Hormones
T4
T3
rT3
Actions of T3 and T4.
T3 and T4 regulate growth and development, especially
critical for the fetus, with requirements lasting past the time of
birth.
T3 and T4 impact upon numerous systems including muscle,
bones, CNS development, myelination, dendritic formation,
and synapse formation.
T3 and T4 act as metabolic stimulants (calorigenic effects) to
increase the rate of oxygen consumption by the heart, skeletal
muscle, liver, and kidney.
In adults, brain, spleen, and gonad metabolism is less
susceptible to the effects of T3 and T4. CNS input through the
hypothalamus (TRH) in cold temperatures increase T3 and T4
secretion to increase metabolic rates. T4 and T3 have
cardiovascular effects, increasing the heart rate and force of
contraction to achieve increased cardiac output.
Biosynthesis of thyroid hormones
The thyroid hormones are synthesized in the
follicular cells of the thyroid
The first step to hormone synthesis is the import of
iodide into the follicular cells a process which
depends an ATP-dependent pump which pumps K+ in
and Na+ out of the cell.
The iodide itself is pumped in with Na+ by a
cotransporter also callad sodium/iodide symporter
Active transport of iodide is required into the thyroid
cell since there is a greater than 50 fold higher
concentration of iodide inside the follicular cell than
outside.
Biosynthesis of thyroid hormones
Once in the follicular cell, iodide is converted into iodide by the enzyme thyroid
peroxidase, which uses hydrogen peroxide (H2O2) as a cofactor.
Thyroid peroxidase catalyzes the incorporation of iodide molecule onto both the 3
and/or 5 positions of the phenol rings of tyrosines found in a glycoprotein called
thyroglobulin.
Thyroglobulin has a molecular weight of about 660,000.
Thyroid peroxidase also appears to couple iodinated tyrosine rings to iodinated
phenol rings which it obtains from other iodinated tyrosine residues within the
protein.
Thyroglobulin contains approximately 300 carbohydrate residues and 5500 amino
acids, including 140 tyrosine residues.
Of these 140 tyrosines, but only two to five of these tyrosines are converted into
either T4 or T3! The entire thyroglobulin protein with its thyroid hormones is stored
in the lumen of the thyroid follicle cell.
Thyroid stimulatory hormone (TSH) stimulates enzymatic degradation of
thyroglobulin to effect release of the thyroid hormones.
T4 is formed exclusively in the thyroid and secreted. In contrast, 80% of T3 found
circulating in the blood is derived from metabolism of T4 in peripheral tissues,
especially the liver and kidney
Biosynthesis of thyroid hormones
.
Thyroid hormone receptors
Thyroid hormone receptors occur in the cytoplasm and nucleus.
The thyroid hormones appear to modulate the dimerization of two
proteins which, when dimerized properly, turn on the transcription of
genes which presumably encode for proteins which mediate the
effects observed for the thyroid hormones.
As part of a negative feedback mechanism, the anterior pituitary has
one of the highest densities of thyroid hormone receptors. T4 and T3
cause a decrease in the levels of TSH.
Heart and skeletal muscle also have high amounts of thyroid hormone
receptors, though there are few in the gonads.
The thyroid hormone receptor in the nucleus is a nonbasic protein
with a MW of 60-65,000. The receptor, influenced by the bound
hormone, binds to DNA and regulates mRNA transcription.
Agonists
Agonists are used to treat hypothyroidism (i.e.
myxedema, infant cretinism, or simple goiter).
T4 and T3 are used to replace natural production of
the hormones if loss is due to disease or medical
intervention.
T4 has a 7 day plasma half-life while T3 has a 1.3 day
half-life.
Since most of T3 comes from T4 through metabolism
in the peripheral tissues, particularly the liver (i.e.
thyroid independent T3 formation),
T4 is often prescribed because of its longer half-life.
Liver dysfunction can effect T3 levels. T3 is
considered the active thyroid hormone.
Inhibitors
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ELISA (Enzyme-Linked Immunosorbent
Assay)
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ELISA