Topic 5 - Cell Signalling 2 Flashcards
Describe the speed and time course of contact-dependent signaling
Fast - less than a second to minutes
Describe the speed and timescale of signalling where molecules are relayed through the bloodstream
Slow - minutes to hours
Slow signalling typically involves altering…
Gene transcription
Fast signalling typically involves altering…
Specific proteins via phosphorylation, allosteric changes in ion channels, or actin rearrangements for example
Describe the difference between primary and secondary responses to a signal of a cell
Primary response: Signal induces transcription of a functional protein which acts to change cell behaviour
Secondary: Signal induces transcription of a transcription factor (protein 1) which then induces the production of a second protein which will exert effects causing a change in cell behaviour
What are 4 key ways in which cells are equipped to react to the same ligand (e.g. ACh) in different ways?
- Different receptors present
- Different intracellular signaling proteins present
- Different effector proteins present
- Different genes activated
In cells with the same intracellular machinery, how are they able to react to a signal in different ways?
Some ligands/factors are area-specific and create GRADIENTS, so the cell reacts to different concentrations of the molecule in different ways
Gradients of morphogens in animals are particularly important during…
Development
What are the 2 ways a morphogen can have a concentration gradient?
- Morphogen is area-specific
2. Morphogen is uniformly distributed but an antagonist for the morphogen is area-specific
Large intracellular signalling molecules can be… (kind of molecule)
Proteins exclusively (with very few exceptions)
What is the purpose of a scaffold protein?
Arranges proteins in a signal cascade in the correct order for succinct signalling
Integration of 2 signalling pathways requires a molecule which is activated by…
Coincidence detection
What is the purpose of anchoring in a signal pathway?
Anchors signalling molecules so the signal is localized
(1) and (2) are examples of modulators in a signal transduction pathway
Activators and inhibitors
Molecular switches have two conformations, which are…
Active and inactive
Monomeric GTPases are activated by…
GTP-binding induced by GEF
What are the 2 kinds of common molecular switches? (I.e. modes of “switching”)
Phosphorylation
GTP-hydrolysis
How many protein kinases are there in the human genome?
520
How many protein phosphatases are there in the human genome?
150
What is the difference between a phosphatase and a kinase?
Kinase = phosphorylates Phosphatase = removes phosphate groups
What are the 2 classes of molecules that signal using GTP binding?
Large trimeric g-proteins
Monomeric GTPases
What are 2 common kinds of kinases?
- Serine/threonine kinases
2. Tyrosine kinases
Scaffold proteins serve to minimize (1) and increase (2) of signalling pathways
- Cross talk
2. Specificity
What is the disadvantage of a pathway using a scaffold protein?
Not a lot of opportunity for amplification of the signal
Docking sites on a scaffold proteins are often created by…
Phosphorylating the scaffold
Describe how PIPs can be used as docking sites in signalling pathways
PIPs can be phosphorylated by receptors when they are activated, this attracts the binding of intracellular signalling proteins
What is a major purpose of a scaffold protein other than localizing things in the correct order?
Protein signals have to be in the right order but also in the right ORIENTATION, scaffolds maintain proper orientation so the proteins can interact
If a scaffold is modified to contain a flexible linker, what happens?
The scaffold is less effective, it is less able to orient signal molecules in the correct directions
Describe the response of a signalling pathway in which the curve of concentration:response is sigmoidal
After a certain threshold concentration of a molecule is reached, the response is amplified
Define desensitization
Prolonged exposure to a stimulus decreases a cell’s response to the stimulus
What is the largest family of cell surface receptors?
GPCRs
What kind of pathways do GPCRs activate?
Sensory - sight, smell, taste
How many transmembrane regions are there in a GPCR?
7
What percentage of modern drugs activate or affect GPCRs?
50%
What domains are present on the alpha subunit of a G-protein?
Ras domain (GTPase activity) and AH domain
What turns off the enzyme activity of a G-protein?
The alpha subunit binding to a regulator of G-protein signalling (RGS)
Which of the 3 subunits of a G-protein are lipid-anchored to the membrane?
Alpha and gamma
Which subunit of the G-protein heterotrimer binds GDP/GTP
Alpha
Which 2 of the G-protein heterotrimer subunits tend to act together as a single signalling factor?
Beta and gamma
Rapid response in signalling requires rapid… (2)
- Synthesis
2. Breakdown
How can the activity of cAMP be increased in the cell?
By stimulating a Gs heterotrimer
Describe the subunits of PKA
2 regulatory subunits, 2 catalytic subunits
CREB only binds to its response element once…
It has been phosphorylated
CREB requires both phosphorylation and association of (1) for activation
a transcriptional coactivator, CREB-binding protein
Pi 4,5-bisphosphate is cleaved into… (2)
Diacylglycerol and IP3
What cleaves PI 4,5-bisphosphate into its 2 components?
A Gq subunit
Diacylglycerol goes on to activate…
Protein kinase C
IP3 goes on to…
Enduce the release of Ca++ from the sarcoplasmic reticulum
To inhibit increase in Ca++ in the cytoplasm, it is useful to…
Phosphorylate IP3 into IP4 or IP2
IP3 is the only inositide that can bind to calcium channels
The most important calcium binding protein is…
Calmodulin
What makes calmodulin highly adaptable so that it can bind many types of proteins of different shapes?
Has a “cable” region which allows it to bend and wrap around other proteins
What are the 2 kinds of domains that make up a CamKII complex?
Hub domains and kinase domains
What are the states of CamKII? (4)
Active
Inactive
Partially active
Ca-independent, 50-80% active
With one kinase domain active, CamKII is in the…
Partially active state
With 2 kinase domains activated, CamKII is in the (1) state
Active
With no kinase domains active, but CamKII is still phosphorylating, what state is the complex in?
The Ca++-independent state
CamKII is involved in the important process of (1) in vertebrates
Learning and memory
Describe the effects of ACh on heart muscle cell contractions (pathway)
ACh binds and activates Gi protein > alpha subunit inhibits AC > beta-gamma subunit opens K+ channels
Why is it important to open K+ channels to reduce strength of contractions of heart muscles?
Because if K+ is high in the cell it is more difficult to depolarize it
What are the three ways in which GPCRs can become desensitized?
- Receptors inactivated
- Receptors sequestered
- Receptors downregulated
What is a GRK?
GPCR kinase
What are 2 examples of GRKs?
PKA and PKC
What molecule binds to a GPCR which will eventually lead to signal desensitization?
Arrestin
Do GPCRs and enzyme-coupled cell surface receptors always activate the same signalling pathways?
No - sometimes lead to the same pathway, a lot of cross-linkage
How many transmembrane domains are there on an enzyme-coupled cell surface receptor?
1
An example of na enzyme-coupled cell surface receptor with intrinsic kinase activity is this class of receptors
RTKs
How can a conformational change occur in a receptor (like an RTK) with only one transmembrane domain?
The receptors can dimerize
Phosphorylation of enzyme-coupled cell surface receptors forms a transient sort of…
Scaffold
What is special about the activation of epidermal growth factor receptors?
Both subunits of the dimer are phosphorylated but only one is activated
Do epidermal growth factor receptors undergo transautophosphorylation?
No - only one of the dimers does the phosphorylating
Adaptor proteins in RTK activation are composed largely of these 2 kinds of domains
SH2 and SH3
What molecule does Ras activate?
MAPK
The order of the boring naming scheme of the Ras-MAPK pathway is (in order…)
Raf, Mek, Erk
How is cross-talk between parallel MAPK modules avoided?
Scaffolds
RTK can activate phosphoinositide 3-kinase to create…
Docking sites for signalling molecules