Hormonal signal transduction

Hormones

are released as part of the general adaptive response to external or internal stimuli As first step they interact with specific receptors on/in target tissues.

 Hormone

receptor interaction results in generation of intracellular signal that can either regulate the activity of a select set of genes, or affect the activity of specific proteins including enzymes and transporter or channel proteins.

 The

final outcome of the signal may result in protein synthesis, cell growth and replication. Many other signaling molecules using the same mechanisms include; cytokines, growth factors, metabolites

 According

to the location of specific cellular receptor hormones are classified into two groups: Group I interact with an intra cellular receptor Group II with membrane bound receptors

 Group

I hormones (lipophilic hormones): They diffuse through the plasma membrane to encounter intracellular receptors which can be in the cytosol or the nucleus. The hormone receptor complex first undergoes activation reaction.

 Binding

of Glucocorticoids to their receptor in the cytosol results in release of heat shock protein from the receptor and then the complex is transferred to the nucleus The receptor also contains nuclear localization sequences that assist in the translocation from the cytosol to the nucleus. In the nucleus the complex binds with high affinity to DNA sequence (HRE).

The

DNA-bound, liganded receptor serves as a high affinity binding site for one or more coactivator proteins This results in accelerated gene transcription

 The

thyroid hormones and retinoids diffuse across the plasma membrane and reach the nucleus The cognate receptor for these is already bound to HRE (TRE or RARE). The DNA bound receptor in the absence of the specific ligand is bound to corepressor protein and inhibit transcription.

 The

binding of the ligand releases the corepressor and the receptor-hormone complex now binds a coactivator (s) resulting in activation of gene transcription. Depending on the hormone, the tissue this process results in specific proteins needed for metabolic activities or.

Group II hormones (peptides and catecholamines)

They

have no carriers in the blood and they have short half in blood circulation. They initiate cellular responses after binding to the extracellular domain of membrane bound receptors.

 These

hormones mainly modulate activity of existing proteins, but can also affect transcription. Their action is more rapid as compared with group I hormones Intracellular second messenger or signaling pathways can be one of the following:

 The

signaling pathways: 1- protein kinase A pathway 2- protein kinase C pathway 3- protein kinase G pathway 4- tyrosine Kinases 5- Calcium/calmodulin dependent protein kinase CaM pathway 6- ligand-gated and voltage-gated ion channels.

Receptors

using protein kinase A, and C are called G protein-coupled Receptors (GPCR). GTP-binding protein (G protein) links the hormone receptor complex with the effector enzyme eg AC, or PLC.

cAMP dependent PKA pathway

Many

hormones can stimulate synthesis of cAMP from ATP by adenylyl cyclase and others inhibit the synthesis. Stimulatory s(s) or inhibitory (i) Gs and Gi. s i Glucagon, epinephrene, ACTH, FSH, hCG, NSH,TSH, PTH, LH, CRH

 Cholera

toxin and Pertussis toxin catalyse ADP-ribosylation of s and i respectively. In case of as the binding of cholera toxin disrupts the GTP-ase activity, thus a subunit can not reassociate with the beta gamma subunits and result in sustain activity of the G protein. Many families of the G proteins with wide range effects

 cAMP

formed by AC binds PKA which a heterodimer molecule consisting of two regulatory subunits R and catalytic subunits C. cAMP binds the R subunit and results in dissociation of the C which becomes activated.

 The

active C subunit phosphorylates a variety of cellular enzymes on serine and threonine residues at specific sites (R/K-X-S/T and R-K-XX-S). Different effects of cAMP such as glycogenolysis, lipolysis, steroidogenesis, secretion, ion transport, enzyme induction, synaptic transmissionare mediated by phosphorylation of the specific enzymes.

 cAMP

effects on transcription are mediated by the protein cyclic AMP response element binding protein (CREB). It is also active when it is phosphorylated by PKA by binding a coactivator protein.

 The

action of cAMP-dependent enzymes is terminated by hydrolysis of cAMP to 5-AMP by phosphodiesterases (PDE). Inhibitors of PDE such caffeine increase intracellular cAMP and mimic or prolong the actions of hormones through this signal

Protein kinase C pathway

The

membrane glycerophospholipid phosphatidylinositol 4,5 bisphosphate is also involved in hormonal signal transduction. It is hydrolysed to DAG and IP3 by the enzyme phospholipase C

The

receptors for acetylcholine, ADH, angiogenin, GRP, TRHwhen coupled by their respective ligands they activate PLC, which is attached to the inner leaflet of the plasma membrane. They first activate Gq protein which is similar to Gs.

 Hydrolysis

results in two second messengers; Diacylglycerol (DAG) and inositol 1,4,5 triphosphate (IP3). DAG activates protein kinase C (PKC) in the presence of Ca2+. IP3 by interacting with a specific intracellular receptor, is an effective releaser of Ca2+ from intracellular storage (ER).

 PKC

phosphorylates many cellular proteins on serine and threonine residues. There several isoenzymes of PKC in different tissues. The action of a group of compounds known as tumor promoters, such phorbol esters, is attributable to their effects on PKC.

 They

mimic cellular DAG as second messengers, but unlike DAG they are not rapidly metabolized and give sustained action. Lethium as antidepressant

Calcium as second messenger

Calcium

triggers many cellular responses such as exocytosis in neurons and endocrine cells, muscle contraction, cytoskeleton rearrangements. It is normally kept very low in the cytosol < 10-7 M by the action of Ca2+ pump in the ER, mitochondria and plasma membrane

 Hormonal,

neuronal or other signals cause either influx of Ca2+ into the cell through specific Ca2+ channels in the plasma membrane or release of sequestered Ca2+ in ER or mitochondria raising cytosolic calcium.

 Changes

in intracellular Ca2+ are detected by calcium-binding proteins that regulate a variety of calcium dependent enzymes. Calmodulin (CaM; M 17000) is an acidic protein with four high affinity Calcium binding sites. When intracellular calcium rises to about 10-6 M, the binding of Ca to calmodulin drives conformational change.

 Calmodulin

associates with a variety of proteins and, in its calcium-bound state, modulates their activities. Calmodulin is a member of calcium binding proteins that also includes troponin, which triggers skeletal muscle contraction in response to increased calcium

 Calmodulin

is an integral subunit of Ca2+/calmodulin-dependent protein kinase (CaM kinase). When intracellular calcium is increased in response to some stimulus, calmodulin binds calcium, undergoes a change in conformation, and activates CaM kinase. The kinase then phosphorylates a number of target enzymes, regulating their activity (synapsin 1).

 Calmodulin

is also regulatory subunit of phosphorylase b kinase in the muscle which is activated by calcium. Thus, calcium triggers ATPrequiring muscle contraction while also activating glycogen breakdown, providing fuel for ATP synthesis.

Ligand-gated ion channels

cGMP dependent protein kinase G

cGMP

is synthesized GTP by guanylyl cyclase (GC) which exists in soluble and membrane bound form. Atrio-natriuretic Factor (ANF) causes natriuresis, diuresis, vasodilation and inhibition of aldosterone secretion. It acts by binding membrane bound GC which synthesizes cGMP from GTPPKG

 Many

compounds including nitroprusside, nitroglycerin, NO, sodium azide, all cause smooth muscle relaxation and are potent vasodilators. They activate soluble GC. Inhibitors of cGMP PDE enhance and prolong these responses (sildenafil, Viagra).

Tyrosine kinases
EGF,

Insulin, IGF receptors contain intrinsic ligand-activated tyrosine kinase activity. Many receptors involved growth control, differentiation and the inflammatory response have this intrinsic tyrosine kinase activity.

 Ligand

receptor interaction results in a tyrosine phosphorylation event that initiates a cascade which may involve several protein kinases, phosphatases.

Insulin receptor
The

insulin receptor is a heterodimer composed of two identical alpha subunits and two beta subunits. When the hormone is bound it is autophosphorylated on specific tyrosine residues.

 Then,

the phosphorylated receptor phosphorylates IRSs on tyrosine residues, at least 4 IRSs exist. IRS binds the Src homology 2 (SH2) domains of a variety of proteins that are directly involved in mediating different effects of insulin: PI-3 kinase PDK1.

Decreases lipolysis Decreases gluconeogenesis Increases lipogenesis Increase protein synthesis and decreases protein degradation

Hormone-Receptor Adenylate-cyclase (Inactive)

Adenylate-cyclase (active)

Copy Right H. Kh. Atif. 2005

ATP

cAMP

Protein kinase-A Slide 69 R2C2(inactive) FA

Protein kinase-A Hormone 2C(active) + R2-cAMP TAG-lipase

(active)

TAG-lipase
(inactive)

FA FA Phospho-protein Phosphatase (inactive)

TAG