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1D Biochemistry: Intracellular Traffic, Protein Sorting and Biosginaling (Group 2 Trans Team) 1 of 16
Bi
Intracellular Traffic, Protein Sorting and Biosginaling
2. Interacts with TOM 20/22 Macromolecule export from NUCLEUS
3. Transferred to import channel TOM 40 Exportins: similar to importins; involved in export of
4. Translocation across inner mt membrane via a complex with many macromolecules
TIM 23 and 17 Cargo molecules for export carry nuclear export signals
proton-motive force across inner mt membrane is (NESs)
required for transport (electric potential or pH Karyopherins: family of importins and exportins
gradient) Translocation of mRNA molecules
Positively charged leader sequence may be helped Attached to a complex named mRNP exporter
through the membrane by the negative charge in the Ran is not involved
matrix Hydrolyzes ATP by an RNA helicase to drive
5. On the inside of inner mt membrane, it interacts with mt translocation
Hsp70 then interacts with Tim 44
6. Targeting sequence is cleaved by matrix (processing)
protease assumes its final shape (with or without Protein transport to PEROXISOME
chaperones) Its proteins are synthesized on cytosolic polyribosome and
mt Hsp70 ensures proper import into matrix and fold prior to import
prevents misfolding or aggregration Contains peroxisomal-matrix targeting sequences (PTSs)
mt Hsp60-Hsp10 system ensures proper folding o PTS 1: tripeptide located at carboxyl terminal; forms
Close apposition at contact sites between outer and complex with cytosolic receptor protein Pex5
inner membranes is necessary for traNslocation to o PTS 2: 9-AA sequence at N-terminus; forms complex
occur with Pex7
Import system can handle intact oligomers
Protein transport to NUCLEUS
Transport is bidirectional occurs through nuclear pore
complexes (NPCs)
Molecules smaller than 40 kDa pass through NPC via
diffusion
For larger molecules, special translocation mechanisms exists
Proteins carry a nuclear localization signal (NLS) e.g.
Pro2-Lys3-Arg-Lys-Val (rich is basic residues)
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 2 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
ROUGH ENDOPLASMIC RETICULUM BRANCH Movement of Proteins inside cell (Fig 4)
o synthesis in membrane-bound
Proteins sorted via RER branch for various polyribosomes
membranes (ER membrane, Golgi apparatus
o insertion of new proteins from
membrane or plasma membrane) and lysozyme or for
exocytosis. ribosomes into ER membrane or
How does the cell know what proteins are to be lumen from the ribosomes or
processed into sites where they are needed? by transport out of ER
signal peptides (This is SIGNAL HYPOTHESIS in a anterograde transport - cargo in
nutshell) COP II vesicles
o ER to cis-GA
*Properties of Signal Peptides
o Enter Golgi cisternae as
Usually located at N terminal;
mature vesicles
approx. 12-35 AA
retrograde transport - cargo in
Region near N-terminus usually
COP I vesicles
carries positive (+) charge, making
o ER to GA
them attracted to phospholipid
segregation and sorting of
membrane (-)
proteins in trans-Golgi network (exit
AA at cleavage site variable;
side) be carried in either secretory
residues -1 and -3 relative to
or transport vesicles.
cleavage site are small and neutral
Met usually the terminal AA
Has central cluster (hydrophobic;
6-12 AA) and outer cluster
(hydrophilic)
Needed for COTRANSLATIONAL
TRANSLOCATION of secretory
proteins
Secretory/Exocytotic Pathway
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 3 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
o Patients lack almost all lysosomal 5. Polypeptide is released into the lumen of endoplasmic
enzymes, causing lysosomes to reticulum.
contain inclusion bodies
o high levels of plasma lysosomal Posttranslational Translocation
enzymes
enzymes fail to reach their
proper intracellular destination,
and are secreted instead
PROTEIN TRANSLOCATION TO ER
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 4 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
unfolded proteins and ER-ASSOCIATED DEGRADATION (ERAD)
prevent their aggregation.
associated ATPase activity
is seen in their interaction
with unfolded proteins
o Calnexin - calcium-binding ER
membrane protein that binds MHC
antigens and monoglucosylated
glycoproteins, retaining them in ER
until glycoprotein has folded
correctly
o Calreticulin similar to calnexin but
not membrane-bound
Folding enzymes
o protein disulfide isomerase (PDI) -
promotes rapid formation and ER-Associated degradation (ERAD)
reshuffling of disulfide bonds of
proteins until the correct set for Misfolded proteins are disposed through ER-
folding is achieved associated degradation (Figure 7).
o peptidyl prolyl isomerase (PPI) - Selective retrotranslocation of misfolded luminal
accelerates folding of proline- and membrane proteins and ubiquitination
containing proteins by catalyzing happen to target them to proteasomes.
cis-trans isomerization of amino Energy partly supplied by p67, which is an AAA-
acid-proline bonds. ATPase
Possible transmembrane channels - Sec61,
Problems may arise in the proper folding of protein derlin and ERAD e3 ligas, Hrd1 and Doa10, (but
o Misfolded proteins prevent chaperone more evidence still needed to support ths claim)
proteins from being exported to intended
destination UBIQUITINATION
ER stress accumulation of errors; prolonged
interaction of misfolded proteins with chaperone
proteins; activates cell death pathways
CELL RESPONDS through Unfolded Protein
Response
Unfolded Protein Response - transmembrane ER
stress sensors sense levels of misfolded proteins and
initiate signal mechanisms:
o transient inhibition of translation
o production of more chaperones
o increased translation of proteins that
degrade misfolded proteins.
Ubiquitination
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 5 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Ubiquitin - highly conserved, 76 amino-acid protruding extracytoplasmically; IV-multipass membrane
peptide used for tagging proteins for protein
destruction in proteasomes.
Cotranslational Insertion
Two types of protein degradation: 1.) ATP-
requiring (lumabas sa quiz) ubiquitination
(major pathway);and 2.) non-ATP dependent
lysosomal protease
Steps in ubiquitination (Figure 8)
o the C-terminal COO group of
ubiquitin is linked via thioester
bond to the SH group of activating
enzyme E1.
o This activated ubiquitin is
transferred to an SH group of the
conjugating enzyme E2.
o Ligase (E3) then catalyzes the
transfer of ubiquitin from E2 to Cotranslational insertion
epsilon-amino group of the target
protein. Cycles are repeated to Proteins (e.g. LDL receptors-lumabas sa
build up the polyubiquitin chain. quiz) enter ER membrane similar to
o APAT DAPAT! Minimum of four U secretory proteins (Figure 5)
molecules to tag protein for Unlike secretory proteins which are
destruction incorporated into the lumen, these receptors
o Deubiquitinating enzymes in the are retained in the membrane and become
proteasome to cleave U from the transmembrane. Through what?
target protein o highly hydrophobic halt-or stop-
transfer signal.
INCORPORATION OF PROTEINS INTO MEMBRANES The receptor to be inserted most likely exits
the translocon by a lateral gate (Figure 10).
Variations in the way proteins are inserted
Posttranslational Translocation
into membranes are caused by the different
routes of protein insertion. Similar to receptors undergoing cotranslational
four transmembrane protein types insertion, cytosolic proteins (e.g. cytochrome b5) undergoing
Routes of protein insertion these type of insertion enter ER membrane through a lateral
o cotranslational insertion gate.
o posttranslational insertion
Retention in GA followed by retrieval to ER
o retention in the Golgi apparatus
followed by retrieval to the proteins w/ KDEL (Lys-Asp-Glu-Leu) or
endoplasmic reticulum HDEL (His-Asp-Glu-Leu) sequence at their
o retrograde transport from the Golgi carboxyl terminal
apparatus. o They travel first through
anterograde vesicular transport
from ER to GA via vesicles with
coat protein II (COPII)
o In GA, they interact and transiently
stay with KDEL receptor
o They travel back to ER through
retrograde vesicular transport in
vesicles coated with COPI and then
dissociate.
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 6 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Retrograde transport from GA 2 Bud Coat proteins bind to Sar1-GTP. Cargo
Formation proteins become enclosed inside vesicles.
Other proteins (e.g. vesicular components to 3 Pinching off Bud pinches off, forming a completely
be recycled) with non-KDEL-containing coated vesicle. Vesicles move along the
proteins; often w/ C-terminal signal rich in cell through microtubules/actin
filaments.
basic residues.
4 Uncoating The vesicle is uncoated when bound GTP
o Same as in retention in GA
is hydrolyzed to GDP by Sar1
followed by retrieval in ER but 5 Targeting Rab molecules are attached to vesicles
WITHOUT the transient stay in and after switching of Rab-GDP to Rab-GTP.
GA due to interaction with KDEL Tethering A specific GEF Rab effector protein on
receptor target membrane bind to Rab-GTP,
tethering vesicles to target membrane.
TRANSPORT VESICLES 6 Docking v-SNARES pair with cognate t-SNARES
Transport vesicles contain proteins destined for GA or PM. in target membrane to form a four-helix
Non clathrin-containing bundles which docks vesicles and
*Clathrin initiates fusion.
-used in vesicles for exocytosis 7 Fusion When v- and t-SNARES are closely
-consists of three interlocking spirals, w/c interact to aligned, vesicle fuses with membrane and
form a lattice around the vesicle contents are released. GTP is then
- clathrin-free - COPI and II, transport and secretory hydrolyzed to GDP, and the RAB-GDP
vesicles carrying cargo from GA to PM molecules are released into cytosol. An
ATPase (NSF) and -SNAP dissociate
*Steps common to all vesicles: the four-helix bundle so that v- and t-
1. Budding SNARES can be reused.
2. Tethering 8 Recycling Rab and SNARE proteins are taken back
3. Docking to ER to be recycled.
4. Membrane fusion
MEMBRANE ASSEMBLY
*molecular switches
1. Sar formation of COPII vesicles The vesicles that were formed in the previous step will fuse
2. ARF - formation of COPI and clathrin-coated vesicles with the target membrane.
3. Rab inner portion of the vesicle - extracytosolic
side of PM
outer cytoplasmic side of fused vesicle-
remains as the cytoplasmic side of PM
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 7 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
SIGNAL TRANSDUCTION
2. Formation of second messengers via effector proteins
OVERVIEW Effector Proteins signaling proteins distal to but
activated by the agonist-bound receptor
Communication is a basic need in biological processes,
specifically from the induction of embryonic development to o Examples (in G protein mediated pathway)
the integration of physiological responses to environmental - Adenylyl cyclase
changes - Phospholipase
Complex mechanisms are involved to control which signals - Phosphodiesterase
are to be released at what time and what responses should - Ion channels
be elicited as an interpretation of the signal Second Messengers
This communication between cells requires mainly o Generated upon receptor activation and they are
extracellular signal molecules that may either travel over amplified and diffused away from their source to
long or short distances. spread the signal to other parts of the cell
These signals then bind to receptor proteins, which may be o Pass the signal on and bind to other signaling
found on the cell surface or intracellularly. proteins or effector proteins.
The binding activates the receptor and consequently activates o Confer specificity and diversity
one or more molecules (intracellular signaling molecules). The - Same signaling molecule different
processed signals are distributed to effector proteins which response
implement the appropriate change of cell behavior. o Two Types:
This cascade of events involving the recognition of a signal, - Water-soluble (cyclic AMP and Ca2+) diffuse
transmission of the signal and activation of intracellular in the cytosol
events, and response, is called signal transduction. - Lipid soluble (diacylglycerol) diffuse in the
plasma membrane
Major Steps:
1. Recognition of the ligand by the receptor
4 types of ligands or extracellular signaling molecules
o Amines: epinephrine
o Peptides and proteins: angiotensin II and insulin
o Steroids: aldosterone, estrogens, and retinoic
acid
o Small molecules: amino acids, nucleotides, ions,
and gases
Ligands may be agonists or antagonists.
o Agonist - a chemical messenger that binds to a
receptor and elicits a response
o Antagonist - competes for a receptor with a
chemical messenger that is normally present in
the body; deactivates the signal transduction
Ligands may be physiological (naturally occurring) or Second messengers confer specifity.
pharmacological (synthetic molecule)
Two types of receptors:
o Cell surface receptors (for peptides and amines) 3. Transmission of the signal to the appropriate
o Intracellular receptors (for steroid and thyroid intracellular signaling proteins
hormones) help relay the signal into the cell by generating small
intracellular mediators or activating the next effector
protein in the pathway
Scaffold proteins
o bind together groups of interacting signaling
proteins into a complex
o close proximity of the signaling proteins results to
interaction of the components at high local
concentrations and to facilitate fast, efficient and
selective activation
o avoids unwanted cross-talk with other signaling
pathways
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 8 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 9 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
o Paracrine signaling occurs on the immediate or the function of the cell. Important elements of this
local environment thus they are also called Local signaling include a variety of proteins, such as
Mediators. integrins, involved in cell to cell adhesion and
o They are diverse type of signaling molecules like regulation of cell shape and motility
histamine, ATP and adenosine through passive
diffusion.
Autocrine signaling
o Involves the release of a molecule that affects the
same cell or other cells of the same type.
o It is important in tissue growth, organ development,
immune and inflammatory response.
Synaptic signaling
o Occurs in very short distances when neurons
transmit electrical signals along their axon and
release neurotransmitters at synapses that affect the
function of other neurons or cells that are distant
from the neuron cell body.
o The physical relationship between the nerve
terminal and the target cell ensures that the
neurotransmitter is delivered to a specific cell.
Endocrine signals are hormones that are secreted
into the blood and are widely dispersed in the body.
o This requires high affinity and selectivity of cell
receptors because hormones cannot easily
dissociate
o Contact-Independent Signaling Endocrine signals
are hormones that are secreted into the blood and
are widely dispersed in the body.
o This requires high affinity and selectivity of cell
receptors because hormones cannot easily
dissociate
Endocrine signals
o Hormones that are secreted into the blood and are
widely dispersed in the body.
o This requires high affinity and selectivity of cell
receptors because hormones cannot easily
dissociate
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 10 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Cell Surface Receptors Hence, a catalytic receptor is an integral membrane
protein possessing both enzymatic, catalytic and receptor
Receptors that are embedded in the membranes of cells. cations.
They act in cell signaling by receiving (binding Its most distinguishable characteristics are:
to) extracellular molecules. They are specialized integral o Intracellular domains have protein kinase, protein
membrane proteins that allow communication between phosphatase, protease or nucleotide
the cell and the extracellular space. phosphodiesterase activity
The extracellular molecules may o Regulate long-term cell functions on a time scale of
be hormones, neurotransmitters, cytokines, growth minutes to hours
factors, cell adhesion molecules, or nutrients. o Antagonist are usually large, secreted proteins that
They react with the receptor to induce changes in function as paracrine/endocrine growth or
the metabolism and activity of a cell. In the process differentiation factor.
of signal transduction, ligand binding affects a cascading
chemical change through the cell membrane. Enzyme-linked receptors based on activity
a. Receptor Tyrosine Kinase (RTK) the most
common receptor; receptors for epidermal growth
Ligand-gated Ion Channel Receptor factor (EGF), platelet derived growth factor (PDGF),
insulin and other polypeptide growth factor
b. Serine/Threonine Kinase - e.g. TGF- mutation
Groups of transmembrane ion channel proteins which open
to allow ions such as sodium, potassium, calcium and involved in some familial colon cancer cases
c. Guanylate Cyclase Activity - e.g. Atrial Natriuretic
chloride to pass through the membrane in response to the
binding of a chemical messenger, such as Factor (ANF); involved in blood pressure regulation
Receptor Tyrosine Kinase (RTK)
a neurotransmitter.
Neurotransmitters
o Chemical messengers
o Enable neurotransmission Single pass transmembrane proteins
o They transmit signals across a chemical synapse, Bind polypeptide ligands that induce oligomerization
such as a neuromuscular junction, from one neuron to Play important role in cellular growth, differentiation,
another "target" neuron, muscle cell, or gland cell. metabolism, and motility
Many neurotransmitters are synthesized from simple Includes:
and plentiful precursors such as amino acids, which o epidermal growth factor receptor (EGFR)
are readily available from the diet and only require a
small number of biosynthetic steps for conversion.
o A neurotransmitter can influence the function of a
neuron through a remarkable number of mechanisms.
In its direct actions in influencing a neurons electrical
excitability, however, a neurotransmitter acts in only
one of two ways: excitatory or inhibitory.
Excitatory Neurotransmitters
Influences trans-membrane ion flow to increase the
probability that the cell with which it comes in contact will
produce an action potential.
Acetylcholine, glutamate, ATP and serotonin
Inhibitory Neurotransmitters
Influences trans-membrane ion flow to decrease the
probability that the cell with which it comes in contact will
produce an action potential.
Gamma-aminobutyric acid (GABA) and glycine.
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 11 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
o platelet-derived growth factor receptors
o fibroblast growth factor receptors (FGFRs)
o vascular endothelial growth factor receptors
o Met (hepatocyte growth factor/scatter factor
[HGF/SF] receptor)
o Ephs (ephrin receptors)
o insulin receptor
Structure:
1. Extracellular binding domain
2. Single transmembrane helix
3. Cytoplasmic region containing the enzyme domain
(tyrosine kinase)
Pathway:
1. Binding of ligand to both monomers causes
dimerization (producing cross-linked dimers) Mutation of RTKs
2. Tyrosine kinase activation via cross-phosphorylation
(beta subunit of one phosphorylates the other) Usually causes cancer/uncontrolled cell growth since
3. Activated tyrosine kinases act as attachments for RTKs are mostly associated with growth factors
relay proteins containing SH2 domain.
a. ErbB1 = bladder, breast, kidney, non-small-cell lung
Insulin Receptor a. Cetuximab = antibody preventing ligand-
A2 B2 heterotetramer (instead of a single-chain induced activation of EGF-binding
receptor) b. ErbB2 = codes for HER2
Pathway: a. HER2 usually complexes with other HERs
1. Insulin binds to two alpha subunits for effect, but can also have an effect by
2. Two alpha subunits (from two insulin receptor dimers) itself
closes in to each other b. Trastuzumab = binds to extracellular
3. Two beta subunits will also approach each other domain of HER2 receptor ; results to arrest
4. Beta subunits will cross-phosphorylate, activating the during G1 phase
insulin receptor c. ErbB3 = breast, colon, prostate, stomach
5. Phosphorylated tyrosine residue acts as attachment d. ErbB4 = ovarian granulosa
for Insulin Receptor Substrate-1 (IRS-1)
6. Phospohrylated IRS-1 acts as attachment for
Phosphoinositide 3-kinase (PI 3-kinase) ***Fourth type: Tyrosine Associated Receptors
7. PI 3-kinase migrates to cell membrane and
phosphorylates the membrane lipid PIP2 to PIP3 Do not have intrinsic kinase activity
8. PIP3 activates PIP3-dependent protein kinase (PDK-1) Need to associate with those that have kinase activity
(*not in diagram) (Src family and Janus family (JAK))
9. PDK-1 activates protein kinase b (Akt) Undergo JAK (Janus Kinase) STAT (signal
10. The non-membrane bound Akt freely migrates to: transducer and activator of transcription) signaling
a. Activate glycogen synthase pathway
b. Recruit glucose transporters to membrane Ligands: cytokines (interferons and interleukins),
erythropoietin
Pathway:
1. Ligand binding
2. Receptor dimerization activates JAK phosphorylation
of receptor
3. STAT attaches to receptor
4. JAK once again phosphorylates, this time the STATs
5. Phosphorylated STATs dimerizes
6. STAT dimer travels to nucleus for effect
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 12 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Drugs/Toxins:
G-PROTEIN COUPLED RECEPTORS
1. Cholera toxin
membrane receptors that bind ligands that are light a. From Vibrio cholera
sensitive compounds, hormones, pheromones, b. Keeps Gas (s) GTP-bound
neurotransmitters, etc c. Increases cAMP levels
d. Increases Cl, Na, H2O efflux in intestinal
Components: lumen
1. Extracellular Ligand Receptor 2. Pertussis toxin
2. Alpha helix that crosses the membrane 7 times a. From Bordetella pertussis
3. G Protein b. Keeps Gai (i) ADP-ribosylated
4. *GTP/GDP c. Increases cAMP levels
d. Increases mucus secretion
G-Protein Sub-units
1. Beta Sub-unit SECOND MESSENGERS
2. Gamma Sub-unit Small intracellular mediators that are generated
3. Alpha Sub-unit in large numbers as a response to receptor
a. As (s) = activates adenylyl cyclase activation
b. Ai (i) = inhibits adenylyl cyclase Spreads signals to other parts of the cell by
c. Aq (q) = activates phospholipase C diffusing away from the source by binding to or
d. At (t) = activates phosphodiesterase altering the conformation of selected signaling
and/or effector proteins
Cyclic AMP (cAMP)
Pathway: It is a secondary messenger derived from ATP
1. Ligand binding to GPCR catalyzed by the enzyme adenylyl cyclase
2. GPCR undergoes conformational change It is produced when membrane receptors like G-
3. Alpha subunit (with GDP) will exchange GDP for GTP protein coupled receptors are activated.
with the help of Guanine Nucleotide Exchange Factor Its production is also inhibited by by agonists of
(GEF) adenylate cyclase inhibitory Gi protein coupled
4. Alpha subunit (now a monomer) will dissociate from receptors which activate phosphodiesterase
the beta and gamma subunit (now a dimer) which degrades cAMP.
5. Alpha subunit with the attached GTP will proceed to cAMP relays the signals from the GPCR to
activate a specific enzyme/protein (commonly protein kinase A, which is a cAMP dependent
adenylyl cyclase, phosphodiesterase, or enzyme that phosphorylates and activates
phospholipase C) enzymes like glycogen phosphorylase and
6. ***will continue until GDP is hydrolysed to GDP via G transcription factors that regulate gene
protein GTPase and GTPase-accelarating proteins expression.
(GAPs) -> will result to alpha subunit associating
again with beta and gamma subunit
The -adrenergic pathway is a cAMP dependent
pathway ( as shown above) and here is a
summary of steps of the pathway:
(Lifted from 1D 2017 trans)
1. The binding of epinephrine to a site on
the receptor deep within the membrane
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 13 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
i. Inactive Gs dissociates
from adenylyl cyclase,
rendering the cyclase
inactive
ii. Gs reassociates with the
and subunits
iii. Gs is again available to
interact with a hormone-
bound receptor
5. cAMP activates Protein Kinase A (PKA)
a. catalyzes the phosphorylation of
inactive phosphorylase kinase b
yields the active form
6. Phosphorylation of cellular proteins by PKA
causes the cellular response to epinephrine
7. cAMP is degraded, reversing the activation
of PKA.
a. cAMP has a short half life in this
pathway
b. It is quickly degraded by
a. Promotes a conformational change
phosphodiesterase
in the receptors intracellular
domain that affects its interaction
Some hormones act by inhibiting adenylyl cyclase,
with the second protein in the
lowering cAMP levels and suppressing protein
signal-transduction pathway, a
phosphorylation.
heterotrimeric GTP-binding
o The binding of somatostatin to its receptor
stimulatory protein, on the cytosolic
leads to an activation of an inhibitory G
side of the plasma membrane
protein (Gi) counterbalances the effects of
b. When GTP is boind to Gs, Gs
glucagon
stimulates the production of cAMP
o In adipose tissue, prostaglandin E1 (PGE1)
by adenylyl cyclase in the plasma
inhibits adenylyl cyclase, thus lowering
membrane.
[cAMP] and slowing the mobilization of lipid
i. The function of Gs as a
reserved triggered by epinephrine and
molecular switch
glucagon.
resembles Ras receptor
Olfactory transduction is an example of a cAMP
(another class of G
dependent pathway
proteins)
c. Gs ( subunit) occupied by GTP= o The odorant molecule is the odorant
Gs active molecule, which will stimulate the production
of cAMP
i. Can activate adenylyl
o Which will activate different ion channels on
cyclase
d. Gs with GDP bound= GS inactive the membrane
2. Binding of epinephrine enables the receptor o Which will depolarize the olfactory cell that
will create an action potential that will be
to catalyze the displacement of bound GDP
transmitted to the brain.
to GTP converting Gs to its active form
a. As this occurs, the and subunits
of Gs dissociate from subunit.
3. Gsa, with its bound GTP, moves in the plane
of the membrane from the receptor to a
nearby molecule of adenylyl cyclase.
a. Gs is held to the membrane by a
covalently attached palmitoyl group
4. Adenylyl cyclase catalyzes the synthesis of
cAMP from ATP: The association of active
Gs with adenylyl cyclase stimulates the
cyclase to catalyse cAMP synthesis
cytosolic cAMP
a. This stimulation by Gs is self- Cyclic GMP (cGMP)
limiting It is a secondary messenger derived from GTP and it
b. Gs is GTPase that turns itself off acts much like cAMP in which that It activates
by converting its bound GRP to different protein kinases in response to binding of
GDP peptide hormones to the external cell surface.
Guanylate cyclase synthesizes cGMP from GTP.
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 14 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Guanylyl cyclase has two isoforms: Two intracellular second messengers produced
in the hormone sensitive phosphadityl inositol
1. Membrane-spanning guanylyl system.
cyclases (Activated by their Activation of Gq activates the PIP2-specific PLC
extracellular ligands) (Phospholipase C), which catalyzes the production
of IP3 and DAG from the membrane phospholipid,
Phosphatidylinositol 4,5-biphosphate, or PIP2
a. Atrial Natriuretic Factor (ANF)
- Receptors found in cells of renal
collecting ducts and smooth muscle
Both contribute to the activation of protein kinase C
of blood vessels
+ (PKC)
o increased renal excretion of Na and, o Inositol triphosphate a water soluble
Consequently, of water, driven by the compound, diffuses from the plasma
change in osmotic pressure membrane to the ER, where it binds to
o causes relaxation (vasodilation) of the blood specific IP3-gated Ca2+ channels, causing
vessel (blood flow; blood pressure) them to open and increase cytosolic Ca2+
b. Guanylin 2+
o Receptors in intestinal epithelial cells o Diacylglycerol cooperates with Ca in
- activating PKC
(Regulates Cl secretion in the intestine)
Protein Kinase C phosphorylates
o Target of a type of bacterial endotoxin
different cellular proteins which produce
(produced by E.coli) that triggers diarrhea
the cellular response to the hormone
-
(Cl secretion reabsorption of water)
2. Soluble NO-activated guanylyl cyclase
o Found in many tissues (smooth
muscle of the heart and blood
vessels)
2+
o stimulates ion pumps that expel Ca from
the cytosol
reduces forcefulness of contraction
o Nitric oxide (NO) cross plasma membrane
w/o carrier
The visual photo transduction pathway is mediated by
cGMP.
1. Light stimulation of rhodopsin leads to
activation of G-protein, transducing
2. Activated G-protein activates cGMP
PDE
3. PDE hydrolyzes cGMP reducing its
concentration
4. This leads to the closure of Na channels
that will lead to the depolarization of the
cell which will create an action potential
that will be propagated to the brain.
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 15 of 16
Intracellular Traffic, Protein Sorting and Biosginaling
Calcium (Ca2+)
One of the major secondary messenger
- Related to IP3 and DAG as second messenger
- It also binds to Calmodulin, a protein mediator of many
2+
Ca -stimulated enzymatic reactions
2+
o When Calmodulin binds to Ca , it undergoes a
change in conformation, and activates the
2+
Ca /calmodulin-dependent protein
kinases (CaM kinases) phosphorylates
target enzymes, regulating their activities.
REFERENCES
2017 1D Trans
2019 1A Trans
Alberts B, Johnson A, Lewis J, et al.
Molecular Biology of the Cell. 4th edition. New
York: Garland Science; 2002
1D Biochemistry: Intracellular Transport, Protein Sorting and Biosignaling (Group 2 Trans Team) 16 of 16