Chapter 15 Cellular Signal Transduction

The biochemistry and molecular biology department of CMU

When environment changes:

Monadresponds directly. Multicellular organismssignal through elaborate system of intercellular or intracellular communication and consequently regulate functions of organisms.

Signaling molecule Receptor of target cell Intracellular molecule biological effect

Signal transduction

1 Signaling Molecules

Signaling molecules
Signaling molecules, which are released by signal-producing cells, reach and transfer biological signals to their target cells to initiate specific cellular responses.

 Extracellular molecules Intracellular molecules

1. Extracellular molecules
protein & peptides: Hormone, cytokine AA & its derivatives: Gly, Glu, adrenaline, thyroxine Steroid: Sex Hormone, glucocorticosteroid Fatty acid derivatives: prostaglandin

(1) Paracrine signaling (local chemical mediators) Secreted by common cells. Reach neighboring target cells by passive diffusion. Time of action is short. Such as GF, PG

(2) Endocrine signal

Secreted by endocrine cells. Reach target cells by blood circulation. Time of action is long. Such as insulin, thyroxine, adrenalin

(3) Synaptic signal (neurotransmitters)

Secreted by neuronal cells. Reach another neuron by synaptic gap. Time of action is short. Such as Acetylcholine (Ach), noradrenaline

(4) Gaseous signal

Simple structure, half life is short and active in chemistry . Such as NO, CO.

GAS MOLECULE

(5) Autocrine signal

Act back to their own cells. Such as GF, cytokine, interferon, interleukin.

2. Intracellular molecule
Ca2+ DG, ceramide IP3 cAMP cGMP Ras, JAK, Raf ions lipid derivatives carbohydrate derivatives nucleotides proteins

Second messenger:
Small molecules synthesized in cells in response to an external signal are the second messengers, which are responsible for intracellular signal transduction. Such as Ca2+, DG, Cer, IP3, cAMP, cGMP

Third messengers:
Third messengers are the molecules which transmit message from outside to inside of nucleous or from inside to outside of nucleous, also called DNA binding protein.

Proteins and peptides: Hormones, cytokines Effect by membrane Amino acid derivatives: receptors Catecholamines Fatty acid derivatives: Extracellular molecules Prostaglandins Signal molecules Intracellular molecules Effect by intracellular receptors Steroid hormones, Thyroxine, VD3

cAMP, cGMP, IP3, DG, Ca2+

2 Receptor

Receptor
Receptors are specific membrane proteins, which are able to recognize and bind to corresponding ligand molecules, become activated, and transduce signal to next signaling molecules. Glycoprotein or Lipoprotein

ligand
A small molecule that binds specifically to a larger one; for example, a hormone is the ligand for its specific protein receptor.

 Membrane receptors membrane Glycoprotein Intracellular receptors Cytosol or nuclei DNA binding protein

1. membrane receptors
(1) Ligand-gate ion channels type (cyclic receptor) ligandreceptorion channel open or close

(2) G Protein-Coupled Receptors (serpentine R) 1) 7-helices transmembrane receptor

Oligosaccharide unit

Cytosolic side

2) G protein (Guanylate binding protein)

G protein refers to any protein which binds to GDP or GTP and act as signal transduction. G proteins consist of three different subunits (, , -subunit). -subunit carries GTPase activity,

binding and hydrolysis of GTP.

3) Classes of G protein
Gs sACcAMP Gi iACcAMP Gq q PI-PLCIP3+DAG Go oion channel Gt t cGMP PDEcGMP Rhodopsin

Glucagon -adrenaline s AC ACTH -adrenaline angiotensin acetylcholine(M2 M4) GF release inhibitory factor iAC

Cholera toxin Ribosylation of Arg of G G s -ATP ATPase Gs -ADP

AC
cAMP Cl- + H2O HCO3Cavity of intestine diarrhea

Pertussis toxin i -ADP-ribosylation Gi AC cAMP allergy of histamine

4) Effect proteins of G protein

AC cGMP PDE (phosphodiesterase PLC PLA2 Channel protein

cAMP ATP

5) Pathway of G protein linked receptor

H R G protein Es

secondary messeger Protein kinase Phophorylation of Es or functional protein Biological effect

(3) Single transmembrane -helix receptor

Tyrosine protein kinase Receptor (catalytic receptor) IGF R, EGF R Non tyrosine protein kinase Receptor Growth Hormone R, interferon R

Tyrosine protein kinase Receptor or receptor tyrosine kinase (RTK)

Cys-rich domain

Immunoglobulin -like domain

EGFR

IGF-1R

PDGFR

FGFR

Insulin

Cytosol

Intracellular insulin effects

Non-receptor tyrosine kinase (NRTK)

Domain of Downstream molecules of TPKR

SH2 domain (Scr homology 2 domain) SH3 domain PH domain (pleckstrin homology domain)

(4) Guanylate cyclase (GC) receptor

Membrane receptor ANP Soluble receptor NO, CO

2. Intracellular receptor (transcription regulated receptor)

Intracellular R is trans-acting elememt cis-acting element gene expression Localized in the cytosol and/or in the nucleus. ligand: Steroid H, VD3, Thyroxine

3. Properties of binding of H and R

highly specificity highly affinity saturation reversible binding special function model

4. Control of receptor activity

Phosphorylation or dephosphorylation of R Phospholipid of membrane Enzyme catalyzed hydrolysis G protein regulation

5. Function of receptor
(1) Recognize the special ligand (2) Binding to special ligand (3) Signal transduction biological effect

3 Pathway of Signal Transduction

Signal transduction mediated by membrane receptor

cAMP dependent-protein kinase A pathway cGMP dependent PKG pathway Ca2+ dependent PK pathway Tyrosine protein Kinase pathway NF-B pathway TGF- pathway

1. cAMP dependent-protein kinase A pathway

H R G protein AC cAMP PKA
Phosphorylation of Es or functional proteins

Biological effects

(1) cAMP metabolism

ATP

AC

Mg

2+

cAMP

PDE
Mg
2+

5'-AMP

PPi

H2O

PDE Phosphodiesterase AC Adenylate cyclase

NH2 N O O P O O O P O O O P O H OH O N CH2 O H H H OH N N

ATP

PPi

AC
NH2 N N N N N NH2 N N O CH2 O H H OH H H OH N

cAMP

CH2 O H H H H O OH

PDE H2O

O O P O

5'-AMP

P O

(2) Mechanism of cAMP effect Activate cAMP-dependent protein kinase (PKA).

(3) PKA effect

Phosphorylate specifically Ser/Thr residues in several proteins 1 Regulation of metabolism 2 Regulation of gene expression

hormons: glucagon, epinephrine active AC ATP cAMP active PKA P ADP ATP ADP P glycogen synthase (active) glycogen synthase (inactive) phosphorylase b kinase ATP ADP P phosphorylase b phosphorylase a H2O inactive AC ATP phosphorylase b kinase Pi

inactive PKA

Pi

H2O

Pi protein phosphatase-1 inhibitor-1 (active) P

H2O

inhibitor-1 (inactive)

ATP

CRE : cAMP response element (TGACGTCA) CREB: CRE binding protein

CRE

DNA

Transcription
mRNA
EB CR B CRE CREB

CR EB CREB
PKA

CREB EB CR

CREB
PKA

2. Ca2+ dependent PK pathway

(1) Ca2+ -DAG -dependent PKC pathway
H R G protein IP3 ER PIP2 PLC DG Ca2+ PS PKC Phosphorylation of Es or functional proteins Biological effects

[Ca 2+]i 0.01-1 mol/L 10-7 mol/L [Ca 2+]o 2.5mmol/L 10-3 mol/L 5000~10000

1 Function of DG and IP3

IP3 + Ropen of Ca2 channel [Ca2 ]
DG PKC

PS, Ca2

2 Function of PKC regulation of metabolism PKC Ser/Thr-P of R, enzyme, Protein of Mb. Gene expression

Early response: PKC Trans-acting factor-P Third messenger

Immediate early genes Late response: PKC

Third messenger-P Activate genes Cell proliferation

(2) Ca2 -CaM dependent protein kinase pathway

H R G protein PLC IP3 Ca2+ CaM

CaMK
Phosphorylation of Es or functional proteins Biological effects

Calmodulin (CaM
Ca2 binding protein 4 Ca2 + CaM Ca2 - CaM CaM kinase Ser/Thr - P Ca2 pump, AC GC Es (glycogen synthase, phosphorylase kinase)

3. cGMP-dependent PKG pathway

ANP Receptorlinked GC cGMP PKG
Phosphorylation of Es or functional proteins

NO, CO Soluble GC

Biological effects

(1) cGMP
GTP
GC

Mg

2+

cGMP

PDE
Mg
2+

5'-GMP

PPi

H2O

GC: Guanylate cyclase

(2) Function of PKG Ser/Thr- P of protein and E

peptides GC NO cGMP PKG Vascular dilatation

ANP atrial natriuretic

peptide hormones neurotransmitters G AC ATP cAMP PKA

AFP GC GTP

neurotransmitters hypothalamic pituitrin G PL DG PKC cell memberane PIP2(PC) IP3

cGMP PKG Ca2+

Ca2+-CaM CaM-PK

phosphorylation of enzymes or proteins trans-acting factor cis-acting element expression P

effects

nucleus

4. Tyrosine-protein kinase pathway (TPK)

TPK receptor is related to proliferation, differentiation, dissociation, carcinomatous change. TPK receptor TPK Mb. non receptor TPK cytosol

(1) Receptor TPK Ras MAPK pathway

GRB2, SOS, Ras, Raf Small G protein: Ras MAPK (mitogen-activated protein kinase) MAPK MAPKK MAPKKK

EGF, PDGF receptor TPK TPK

GRB2 P SOS P Ras GTP Raf P MAPKK P MAPK P

nucleus trans-acting factor P

expression

(2) JAKs STAT pathway

ligand non TPK receptor JAKs STAT

gene expression

 JAKs STAT: Signal transductors and activator of transcription

interferons Plasma membrane

JAK

P84 113 91

- P
STAT complex 113 91 84 P Nuclear membrane

P
48

inactive STAT

P
113

P
gene

91
48

84

transcription

Interferons response element

Interferon Dimerization of Receptor Autophosphorylation of JAK Phosphorylation of STAT Nuclear translocation of STAT Interferon response element Expression of gene

5. Nuclear factor-B pathway

6. TGF-pathway

Intracellular receptor (DNA transcription regulated receptor)

Steroid H, VD3, Thyroxine Cytosolic R: glycocorticosteroid H Nuclear R: thyroxine, estrogen, androgen, progesterone

Serum binding protein With bound bormone