SIGNAL TRANSDUCTION AND REGULATION

Sigit Purwantomo spurwantomo@mrinstitute.org

Subject of this weeks lecture

The importance of signal transduction Malfunction on signaling pathways

Weekly objective
Cell signaling and signal transduction

Regulation of the cell cycle

Malfunction of signaling pathways

Lecture 1

Cell signaling & signal transduction

Cell communication
Cell signaling

Signal transduction

Why we need communication?

Pay attention for the next slide!

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Theres something wrong with your phone I cant hear you

The important of communication

Signal

Signal perception Signal transduction Response

Cell

General feature of cell signaling & signal transduction

Blue : terminator signal

Modes of Cell-Cell Signaling

In endocrine signaling, hormones are carried through the circulatory system to act on distant target cells. Cell signaling can take place either through direct cell-cell contacts or through the action of secreted signaling molecules. In paracrine signaling, a molecule released from one cell acts locally to affect nearby target cells.

Endocrine

In autocrine signaling, a cell produces a signaling molecule to which it also responds.

Type of signal transducers

1 3 5

4
6

Gated ion-channel (1)

Na/K ATPase
3 Na = 2 K
Membrane potential

Na K ATPase

Gated Na channel: structure

Consist of 4 domains There are 6 helices in each domain

Helix no. 4 function = voltage sensor Helix no. 6 function = activating gate

Activation gate

Nicotinic acetylcholine receptor: structure

L H

5 subunit

Acetylcholine will bind to alpha

Nicotinic acetylcholine receptor: mechanism

Subunit folds into 4 transmembranes helices

4 helices in each subunit

Nicotinic acetylcholine receptor: in action

Keyword : twisting

hydrophobic

polar

Gate closed (resting)

Gate open (exited)

Gated K channel: structure

4 subunit

Cl channel

Cl channel

How does the chloride channel work?

The chloride channel is made from a protein called CF transmembrane regulator (CFTR) protein. Its normal function is to control the flow of chloride ions from the cell.

How does the chloride channel work?

Phosphate groups (P) add to the R-domain. Cyclic AMP (cAMP) stimulates the enzyme, protein kinase (PKA), to add the phosphate groups.

The channel is closed ATP is bound . . .

The shape of CFTR changes, opening the Clchannel

. . . and hydrolysed to ADP+Pi

Cl channel & cystic fibrosis (CF)

The basics

There are channels in these lining cells through which ions can pass.

Normally, the movements of ions brings water to the surface of the airway and keeps the mucus moist.
The basic defect in CF arises particularly in the epithelial cells lining the airways of the lung
http://resources.schoolscience.co.uk/MRC/3/page3.html

How do the channels keep the mucus moist?

The lining cells have channels on their outside surface (on the side of the airway).

One of the channels allows sodium ions to flow into the cell and the other controls the passage of chloride ions out of the cell into the mucus on the airway surface.

How do the channels keep the mucus moist?

Along with the ion pump, the action of the channels results in an excess of chloride ions in the mucus; i.e. an ionic gradient is set up, with a higher concentration towards the outside.

In an attempt to equalise the salt concentrations, water is dragged out through the gaps between the cells and this keeps the mucus moist.

What happens in CF?

In the lining cell of a person with CF, the vital chloride channel is blocked.

This means that there is no movement of chloride ions into the mucus. With no ionic gradient, there is no need for water to move towards the surface and the mucus dries out.

Type of signal transducers

1 3 5

4
6

Receptor enzyme (2)

Receptor Protein-Tyrosine Kinases

Directly linked to intracellular enzyme Dimerization and autophosphorylation of receptor protein-tyrosine kinases

Growth factor binding induces receptor dimerization, which results in receptor autophosphorylation as the two polypeptide chains phosphorylate one another.

Some examples

EGF : epidermal growth factor PDGF : platelet derived growth factor (blood vessel formation, angiogenesis)

Downstream signaling molecules with receptor protein-tyrosine kinases

SH2 domains bind to specific phosphotyrosinecontaining peptides of the activated receptors. SH2 : Src homology 2 Src (pronounced sarcas it is short for sarcoma)

SH2 : Src homology 2

Insulin receptor
Extracellular

Insulin structure

Insulin biosynthesis

Release of insulin by the b-cells

Glucose enters the cell [ATP]/[ADP] ratio increases, ATP-dependent K channel (K ATP) is closed The closing of this channel leads to a membrane depolarization Ca2 enter the cell such that intracellular Ca2 levels increase

The increase in intracellular Ca2 stimulates insulin secretion

Type of signal transducers

1 3 5

4
6

Protein receptor (3)

G protein coupled-receptor
Characterized by seven transmembrane a helices.

Structure of a G proteincoupled receptor

Regulation of G proteins
4

Hormonal activation of adenylyl cyclase

A guanine nucleotide-binding protein (called a G protein) is an intermediary in adenylyl cyclase activation

cAMP synthesis and degradation

cAMP is synthesized from ATP by adenylyl cyclase

Cyclic AMP is degraded to AMP by cAMP phosphodiesterase.

Cyclic AMP-inducible gene expression

The free catalytic subunit of protein kinase A translocates to the nucleus and phosphorylates the transcription factor CREB (CRE-binding protein), leading to the recruitment of coactivators and expression of cAMP-inducible genes.

1 2 3

Activation protein kinase A Phosphorilation CREB Expression cAMP-inducible genes

CRE : cAMP response element

Regulation of protein kinase A

1
The inactive form of protein kinase A consists of two regulatory (R) and two catalytic (C) subunits.

Binding of cAMP to the regulatory subunits induces dissociation of the catalytic subunits, which are then enzymatically active.

Type of signal transducers

1 3 5

4
6

Steroid receptor (4)

Principles of hormone action

Mechanism of action

The receptor

Type of signal transducers

1 3 5

4
6

Receptor with no enzyme activity (5)

The JAK/STAT pathway

In unstimulated cells, STAT proteins are inactive in the cytosol.

STAT proteins are phosphorylated by the receptor-associated JAK protein-tyrosine kinases. The phosphorylated STAT proteins dimerize and translocate to the nucleus Activation of transcription of target genes.

STAT : signal transducer and activator transcription

Type of signal transducers

1 3 5

4
6

Adhesion receptor (6)

Signal Transduction and the Cytoskeleton