BIOSIGNALING  Open and close in response to the binding of

(from Lehninger) chem. ligands or changes in transmembrane

potential
signal transduction - signal represents information that is detected y  Simplest signal transducers
specific receptors and converted to a cellular response, which always  E.g. acetylcholine receptor ion channel
involves a chemical process 5. Adhesion receptors
 Interact w/ macromolecular components of the
GENERAL FEATURES OF SIGNAL TRANSDUCTION ECM and convey instructions to the cytoskeletal
 Specificity system about cell migration or adherence to
o Achieved by precise molecular complementarity matrix
between the signal and receptor molecules, mediated  E.g. integrins
by the same kinds of weak (noncovalent) forces that 6. Nuclear receptors
mediate enzyme-substrate ad antigen-antibody  Steroid receptors
interactions  Bind specific ligands and alter the rate at w/c
o Specificity is further achieved by the presence of specific genes are transcribed and ranslated into
receptors in certain cell types only cellular proteins
 Affinity
o The receptor detects picomolar concentrations of a
dignal molecule G PROTEIN-COUPLED RECEPTOR
 Cooperativity  3 essential components that define signal transduction through
o In receptor-ligand interactions, results in large changes GPCR:
in receptor activation with small changes in ligand o Plasma membrane with 7 transmembrane helical
concentration segments
 Amplification by enzyme cascades o An effector enzyme in the plasma membrane that
o When an enzyme asso with a signal receptor is generates an intracellular 2nd messenger
acteivated and catalyzes the activation of many molec o Guanosine nucleotide-binding protein (G protein) that
of a 2nd enzyme, each of w/c activates many molec of a activates the effector enzyme
third enzyme, and so on  Mechanism of action
 Desensitization o G protein exchanges bound GDP to GTP
o When the stimulus falls below a certain threshold, the o GTP-protein dissociates from the occupied receptor
system again becomes sensitive and binds to a nearby enzyme, altering its activity
 Integration  Implicated in many common human disease (allergies, epression,
o Ability of system to receive multiple signals and blindness, diabetes, CV defects)
produce a unified response appropriate to the needs  B-adrenergic receptor is the prototype for all GPCRs
of the cell or organism
B-Adrenergic Receptor Systems
 Epineprine - Fight or flight
6 BASIC RECEPTOR TYPES o Epi action begins when hormone binds to a protein
1. G protein-coupled receptor receptor in the plasma membrane of an epi-sensitive
 Indirectly activate enzymes that generate cell
intracellular second messangers  Adrenergic receptors – α1, α2, β1, β2 (has differences in
 e.g. B-adrenergic receptor (detects epi) affinities and responses to a group of agonists and antagonists
2. Receptor tyrosine kinases o Agonists – structural analogs that bind to a receptor
 Plasma membrane receptors; enzymes and mimic the effects of its natural ligand
 Catalyzes the phosphorylation of several cytosolic o Antagonists – analogs that bind the receptor w/o
or plasma membrane proteins when this receptor trigerring the normal effect and thereby blocking the
is activated by its extracellular ligand effects of agonists including the biological ligand
 E.g. insulin receptor; EGF-R o 4 types are found in diff target tissues and mediate diff
3. Receptor guanylyl cyclases responses to epi
 Plasma membrane receptow w/ an enzymatic  B-adrenergic receptor in muscle, liver and adipose tissue
cytoplasmic domain o Mediate changes in fuel metab (increased breakdown
 cGMP (intracellular 2nd messenger of this of glycogen and fat)
receptor) activates a cytosolic protein kinase that o Integral protein with 7 hydrophobic regions of 20-28
phosphorylates cellular proteins and changes AA residues that snake back and forth across the
their activities plasma membrane 7 times; “serpentine receptors” or
4. Gated ion channels “heptahelical receptors”
o Steps:  Association of active Gsα qith adenylyl cyclase
 Epi binds to a receptor deep w/in the plasma stimulate cyclase ro catalyze cAMP synthesis
membrane from ATP
 Conformational change in the receptor’s  Possible only when switch regions
intracellular domain w/c affects its of Gsα are exposed by a GTP-
interaction with the 2nd protein, Gs, in signal induced conformational change
transduction pathway on the cytoplasmic o Gs can turn itself off
side of membrane  Convert bound GTp to GDP
 active Gs stimulates production of  Inactive Gsα dissociates from adenylyl cyclase,
cAMP by adenylyl cyclase in plasma therefore cyclase is inactive
membrane  Gsα reassociates w/ βγ dimer
 when α-subunit (nucleotide-  Inactive Gsα again available to interact with
binding site of Gs) is occupied by hormone-bound receptor
GTP, Gs is turned on (if GDP is o Inactive form of PKA contains 2 identical catalytic
bound to site, Gs is off) subunits and 2 identical regulatory subunits
 Gs can activate adeylyl cyclase  R2C2 complex is inactive.
 activated B-adrenergic receptor  Autoinhibitory domain of each R subunit
interacts with Gs, catalyzing occupies substrate-binding cleft of each C
replacement of bound GDP with subunit
GTP and converting Gs to its active  Conformational change that moves
form autoinhibitory domain of R out of C
 β and γ subunits of Gs dissociate  R2C2 complex dissociates
from α subunit as βγ dimer  Yield 2 free, catalytically active C subunits
 Gsα is held to the membrane by a
covalently attached palmitoyl Termination of B-adrenergic response
group 1. Desensitization – mediated by protein kinase that
 Activation of adenylyl cyclase = phosphorylates the receptor in the intracellular domain that
increase in cAMP normally interacts with Gs
 cAMP activates cAMP-dependent a. Binding of Epi to B-adrenergic receptor triggers
protein kinase (PKA) which dissociation of Gsβγ from Gsα
catalyzes phosphorylation of other b. Gsβγ recruits βARK to the membrane, where it
proteins (like PKB that when phosphorylates Ser residues at the carboxyl terminus
phosphorylated, mobilizes of receptor
glycogen stores in muscle and liver  BARK is a member of family of G protein
to glucose for energy) coupled receptor kinases (GRKs), all of w/c
o Steps: (in short. :D) phosphorylate GPCRs on their carboxyl-
 Epi binds to its specific receptor terminal cytoplasmic domains and play roles
 The occupied receptor causes replacement similar to that of βARK in desensitization and
of the GDP bound to Gs by GTP, activation resensitization of receptors
Gs. c. Β-arrestin (βarr) binds to the phosphorylated carboxyl-
 Gs (α-subunit) moves to adenylyl cyclase and terminal domain of receptor
activates it. d. Receptor-arrestin complex enters cell by endocytosis
 Stimulation of Gs is self-limiting e. In endocytic vesicle, arrestin dissociates; receptor is
 Adenylyl cyclase catalyzes the formation of dephosphorylated and returned to cell surface
cAMP. 2. Hydrolysis of GTP bound to the Gα subunit of the intrinsic
 cAMP activates PKA. GTPase activity of G protein
 Phosphorylation of cellular proteins by PKA a. GTP to GDP = return Gα to conformation with βγ
causes the cellular response to epinephrine. subunits = G protein unable to interact w/ adenylyl
 cAMP is degraded by cyclic nucleotide cyclase = end production of cAMP
phosphodiesterase to 5’-AMP, reversing the b. GTPase activator protein (GAPs) stimulate GTPase
activayion of PKA. activity = rapid inactivation of G protein
o Adenyl cyclase 3. Remove 2nd messenger
 Integral protein of plasma membrane; active a. Hydrolysis of cAMP to 5’-AMP by cyclic mucleotide
site on the cytoplasmic face phosphodiesterase

cAMP as 2nd messenger for many regulatory molecules

 Epi change the intracellular cAMP, thus, change activity of PKA  Ca++ level does not simply rise and then decrase, but rather
 Catalytic subunit of PKA can also move into the nucleus where it oscillates with a period of a few seconds
phosphorylates the cAMP response element binding protein  In some tissues, adenylyl cyclase that produces cAMP, and
(CREB) which alters the expression of specific genes regulated by phosphodiesterase that degrades cAMP are stimulated by Ca++
cAMP o Temporal and spatial changes in Ca++ can produce
 Inhibit adenylyl cyclase, lower cAMP, suppress protein transient, localized changes in cAMP
phosphorylation (summary: page 439; Lehninger)
o Binding of somatostatin (counterbalance the effects of
glucagon) to receptor leads to activation of inhibitory RECEPTOR TYROSINE KINASES
G protein  Large family of pasma membrane receptors with intrinsic protein
 Many types of signals can be mediated by cAMP because of the kinase activity
confinement of the signaling process to a specific ergion of the  Have ligand-binding domain on the extracellular face of the
cell by adaptor proteins (noncatalytic proteins that hold together plasma membrane and an enzyme active site on the cytoplasmic
other protein molec that fxn in concert) face connected by a single transmembrane segment
o AKAPs (A kinase anchoring proteins) – multivalent o Cytoplasmic domain: protein kinase that
adaptor proteins; one part bind to R subunit of PKA phosphorylates Tyr residues in specific target protein
 Receptors for insulin and epidermal growth factor
Related roles of DAG, ITP and Ca++ as 2nd messengers
 A second broad class of GPCRs are coupled through a G protein Stimulation of Insulin Receptor
to a plasma membrane phospholipase C (PLC) that is specific for  Insulin regulates both metabolic enzymes and gene expression
the membrane phospholipid phosphatidylinositol 4,5-  Insulin does not enter cells; initiates signal that travels a
bisphosphate (PIP2) branched pathway from the plasma membrane receptor to
1. Hormone binds to specific receptor insulin-sensitive enzymes in the cytosol and nucleus
2. The occupied receptor causes GDP-GTP exchange on  Steps:
associated G protein (Gq) o Insulin receptor (INS-R) binds insulin and undergoes
3. Gq, with bound GTP, moves to PLC and activates it autophosphorylation on its carboxyl-terminal Tyr
4. Active PLC cleaves PIP2 to inositol triphosphate (IP3) residues
and DAG  INS-R has α subunits (contain insulin-binding
5. IP3 binds to a specific receptor on the ER, releasing domain) protruding from the outer face of
sequestered Ca++ the plasma membrane. Also has 2
6. DAG and Ca++ activate PKC at the surface of plasma transmembrane Β subunits (protein kinase
membrane activity) with carboxyl termini protruding
7. Phosphorylation of cellular proteins by PKC produces into the cytosol.
some of the cellular responses to the hormone  INS-R is already an (αβ)2 dimer before insulin
binds.
Ca is a 2nd messenger that may be localized in space and time o Insulin receptor phosphorylates IRS-1 (Insulin receptor
 Ca++ serves as 2nd messenger that triggers intracellular substrate-1) on its Tyr residues
responses  IRS-1 becomes point of nucleation for
o Exocytosis in neuron and endocrine cells complex of proteins that carry the message
o Contraction in muscles from the insulin receptor to end targets in
o Cytoskeletal arrgts during ameboid movement the cytosol and nucleus, through long series
 Cytosolic Ca++ is kept very low in unstimulated cells of intermediate proteins.
 Hormonal, neural or other stimuli case either influx of Ca++ into o SH2 (Src homology 2) domain of protein Grb2 binds to
cell ot the release of sequestered Ca++ from ER or mitochondria phosphorylated Tyr of IRS-1. Sos binds to SH3 of Grb2,
 Chanes in intracellular Ca++ are detected by Ca++-binding then to Ras, causing GDP release and GTP binding to
proteins that regulate a variety of Ca++-dependent enzymes Ras.
 Calmodulin – acidic protein with 4 high-affinity Ca++-binding site  Grb2 is an adaptor protein, with no intrinsic
o Binding of Ca++ to calmodulin drives conformational enzymatic activity; fxn is to bring together 2
change when intracellular Ca++ rises; thus, activates proteins that must interact to enable signal
CaM kinase transduction.
o Integral subunit of the Ca++/calmodulin-dependent  Sos acts as a guanosine nucleotide-exchange
protein kinases (CaM kinases) factor (GEF), catalyzing the replacement of
 Activity is spatially restrictd; after its release triggers a local bound GDP with GTP on Ras, a G protein
response, Ca++ is generally removed before it can diffuse to  Ras – prototype of family of small G proteins;
distant parts of the cell can exist in either GTP-bound or GDP-bound
conformation
 o Activated Ras binds and activates Raf-1.
o Raf-1 phosphorylates MEK on two Ser residues,
activating it. MEK phosphorylates ERK on a Thr and a
Tyr residue, activating it.
o ERK moves into the nucleus and phosphorylates
nuclear transcription factors such as Elk1, activating
them.
o Phosphorylated Elk1 joins SRF to stiulate the
transcription and translation of a set of genes needed
for cell division.
 100 insulin-related genes
 ERK is in MAPK family (mitogen-activated protein kinases)
 Kinases in the MAPK and MAPKKK families are specific to Ser or
Thr residues
 MAPKK phosphorylate both a Ser and a Tyr residues in their
substrate, MAPK (here, ERK)
 MAPK cascades – mediate signaling initiated by a variety of GF

Membrane Phospholipid PIP3

 Enzyme phosphoiositide-3 kinase also binds to IRS-1 (not only
Grb2 binds to IRS-1)
 Steps
o IRS-1, phosphorylated by insulin receptor, activates PI-
3K by binding to its SH2 domain. PI-3K converts PIP2 to
PIP3.
o PKB bound to PIP3 is phosphorylated by PDK1. Thus
activated, PKB phosphorylates GSK3 on a Ser residue,
inactivating it.
o GSK3, inactivated by phosphorylation, cannot convert
glycogen synthase (GS) to its inactive form by
phosphorylation, so GS remains active.
o Synthesis of glycogen from glucose is accelerated.
o PKB stimulate movement of GLUT4 from internal
membrane vesicles to the plasma membrane,
increasing the uptake of glucose.

JAK-STAT Signaling System

 When EPO binds to its plasma membrane receptor, the receptor
dimerizes, and the dimer can bind and activate the soluble
protein kinase JAK (JAK kinase).
 Activated JAK phosphorylates several Tyr residues in the
cytoplasmic domain of the EPO receptor. STATs (signal
transducers and activators of transcription) are also targets of
JAK.
 An SH2 domain in STAT5 binds phosphorylated Tyr residues in
the EPO receptor, positioning the STAT for phosphorylation by
JAK in response to EPO.
 PHosphorylated STAT5 forms dimmers, exposing a signal that
causes it to be transported in the nucleus.
 In nucleus, STAT5 induces the transcription of specific genes
essential for erythrocyte maturation.

*Src – first protein found to have the char phosphorylated-Tyr-

binding domain (SH2 domain).

(Summary: page 444, Lehninger)