Signal relay pathways
Learn how signals are relayed inside a cell starting from the cell membrane receptor. The chains of molecules that relay intracellular signals are known as intracellular signal transduction pathways.
Once a signaling molecule (ligand) from one cell has bound to a receptor on another cell, is the signaling process complete?
If we're talking about intracellular receptors, which bind their ligand inside of the cell and directly activate genes, the answer may be yes. In most cases, though, the answer is no—not by a long shot! For receptors located on the cell membrane, the signal must be passed on through other molecules in the cell, in a sort of cellular game of "telephone."
The chains of molecules that relay signals inside a cell are known as intracellular signal transduction pathways. Here, we’ll look at the general characteristics of intracellular signal transduction pathways, as well as some relay mechanisms commonly used in these pathways.
Binding initiates a signaling pathway
When a ligand binds to a cell-surface receptor, the receptor’s intracellular domain (part inside the cell) changes in some way. Generally, it takes on a new shape, which may make it active as an enzyme or let it bind other molecules.
The change in the receptor sets off a series of signaling events. For instance, the receptor may turn on another signaling molecule inside of the cell, which in turn activates its own target. This chain reaction can eventually lead to a change in the cell's behavior or characteristics, as shown in the cartoon below.
Cartoon-style schematic showing how the components of a hypothetical signaling pathway are activated sequentially, with one turning on the next to produce a cellular response.
Because of the directional flow of information, the term upstream is often used to describe molecules and events that come earlier in the relay chain, while downstream may be used to describe those that come later (relative to a particular molecule of interest). For instance, in the diagram, the receptor is downstream of the ligand but upstream of the the proteins in the cytosol. Many signal transduction pathways amplify the initial signal, so that one molecule of ligand can lead to the activation of many molecules of a downstream target.
The molecules that relay a signal are often proteins. However, non-protein molecules like ions and phospholipids can also play important roles.
The cartoon above features a bunch of blobs (signaling molecules) labeled as “on” or “off.” What does it actually mean for a blob to be on or off? Proteins can be activated or inactivated in a variety of ways. However, one of the most common tricks for altering protein activity is the addition of a phosphate group to one or more sites on the protein, a process called phosphorylation.
Diagram of a phosphorylated protein bearing a phosphate group attached to a serine residue, showing the actual chemical structure of the linkage.
Phosphate groups can’t be attached to just any part of a protein. Instead, they are typically linked to one of the three amino acids that have hydroxyl (-OH) groups in their side chains: tyrosine, threonine, and serine. The transfer of the phosphate group is catalyzed by an enzyme called a kinase, and cells contain many different kinases that phosphorylate different targets.
Phosphorylation often acts as a switch, but its effects vary among proteins. Sometimes, phosphorylation will make a protein more active (for instance, increasing catalysis or letting it bind to a partner). In other cases, phosphorylation may inactivate the protein or cause it to be broken down.
In general, phosphorylation isn’t permanent. To flip proteins back into their non-phosphorylated state, cells have enzymes called phosphatases, which remove a phosphate group from their targets.
Cartoon-style diagram showing how a protein is phosphorylated by a kinase through the addition of a phosphate from ATP, producing ADP as a by-product, and dephosphorylated by a phosphatase, releasing Pi (inorganic phosphate) as a by-product. The two reactions make up a cycle in which the protein toggles between two states.
Phosphorylation example: MAPK signaling cascade
To get a better sense of how phosphorylation works, let’s examine a real-life example of a signaling pathway that uses this technique: growth factor signaling. Specifically, we'll look at part of the epidermal growth factor (EGF) pathway that acts through a series of kinases to produce a cellular response.
This diagram shows part of the epidermal growth factor signaling pathway:
Phosphorylation (marked as a P) is important at many stages of this pathway.
- When growth factor ligands bind to their receptors, the receptors pair up and act as kinases, attaching phosphate groups to one another’s intracellular tails. Read more in the article on receptors and ligands.
- The activated receptors trigger a series of events (skipped here because they don't involve phosphorylation). These events activate the kinase Raf.
- Active Raf phosphorylates and activates MEK, which phosphorylates and activates the ERKs.
- The ERKs phosphorylate and activate a variety of target molecules. These include transcription factors, like c-Myc, as well as cytoplasmic targets. The activated targets promote cell growth and division.
Together, Raf, MEK, and the ERKs make up a three-tiered kinase signaling pathway called a mitogen-activated protein kinase (MAPK) cascade. (A mitogen is a signal that causes cells to undergo mitosis, or divide.) Because they play a central role in promoting cell division, the genes encoding the growth factor receptor, Raf, and c-Myc are all proto-oncogenes, meaning that overactive forms of these proteins are associated with cancer.
MAP kinase signaling pathways are widespread in biology: they are found in a wide range of organisms, from humans to yeast to plants. The similarity of MAPK cascades in diverse organisms suggests that this pathway emerged early in the evolutionary history of life and was already present in a common ancestor of modern-day animals, plants, and fungi.
Although proteins are important in signal transduction pathways, other types of molecules can participate as well. Many pathways involve second messengers, small, non-protein molecules that pass along a signal initiated by the binding of a ligand (the “first messenger”) to its receptor.
Second messengers include ions; cyclic AMP (cAMP), a derivative of ATP; and inositol phosphates, which are made from phospholipids.
Calcium ions are a widely used type of second messenger. In most cells, the concentration of calcium ions () in the cytosol is very low, as ion pumps in the plasma membrane continually work to remove it. For signaling purposes, may be stored in compartments such as the endoplasmic reticulum.
In pathways that use calcium ions as a second messenger, upstream signaling events release a ligand that binds to and opens ligand-gated calcium ion channels. These channels open and allow the higher levels of that are present outside the cell (or in intracellular storage compartments) to flow into the cytoplasm, raising the concentration of cytoplasmic .
How does the released help pass along the signal? Some proteins in the cell have binding sites for ions, and the released ions attach to these proteins and change their shape (and thus, their activity). The proteins present and the response produced are different in different types of cells. For instance, signaling in the β-cells of the pancreas leads to the release of insulin, while signaling in muscle cells leads to muscle contraction.
Cyclic AMP (cAMP)
Another second messenger used in many different cell types is cyclic adenosine monophosphate (cyclic AMP or cAMP), a small molecule made from ATP. In response to signals, an enzyme called adenylyl cyclase converts ATP into cAMP, removing two phosphates and linking the remaining phosphate to the sugar in a ring shape.
Once generated, cAMP can activate an enzyme called protein kinase A (PKA), enabling it to phosphorylate its targets and pass along the signal. Protein kinase A is found in a variety of types of cells, and it has different target proteins in each. This allows the same cAMP second messenger to produce different responses in different contexts.
Diagram of a pathway that uses cAMP as a second messenger. A ligand binds to a receptor, leading indirectly to activation of adenylyl cyclase, which converts ATP to cAMP. cAMP binds to protein kinase A and activates it, allowing PKA to phosphorylate downstream factors to produce a cellular response.
cAMP signaling is turned off by enzymes called phosphodiesterases, which break the ring of cAMP and turn it into adenosine monophosphate (AMP).
Although we usually think of plasma membrane phospholipids as structural components of the cell, they can also be important participants in signaling. Phospholipids called phosphatidylinositols can be phosphorylated and snipped in half, releasing two fragments that both act as second messengers.
One lipid in this group that's particularly important in signaling is called . In response to a signal, an enzyme called phospholipase C cleaves (chops) into two fragments, DAG and . These fragments made can both act as second messengers.
DAG stays in the plasma membrane and can activate a target called protein kinase C (PKC), allowing it to phosphorylate its own targets. diffuses into the cytoplasm and can bind to ligand-gated calcium channels in the endoplasmic reticulum, releasing that continues the signal cascade.
Image of a signaling pathway that uses inositol triphosphate and calcium ions as second messengers. After a ligand binds to a receptor at the membrane, phospholipase C is indirectly activated. It cleaves PIP2 to produce IP3 and DAG. DAG stays in the membrane and activates protein kinase C, which phosphorylates its targets. The IP3 is released into the cytosol and binds to a calcium ion channel in the endoplasmic reticulum, causing the channel to open. Calcium ions stored in the endoplasmic reticulum rush into the cytosol, where they bind to calcium-binding proteins. The calcium-binding proteins trigger a cellular response.
And...it's even more complicated than that!
Signaling pathways can get very complicated very quickly. For instance, the full version of the epidermal growth factor signaling pathway we saw earlier looks like a huge hairball and takes up an entire poster if you try to draw it out! You can see this for yourself in Sal's video on the MAPK pathway.
This complexity arises because pathways can, and often do, interact with other pathways. When pathways interact, they basically allow the cell to perform logic operations and "calculate" the best response to multiple sources of information. For instance, signals from two different pathways may be needed to activate a response, which is like a logical "AND." Alternatively, either of two pathways may trigger the same response, which is like a logical "OR."
Left diagram: logical "AND" in a cell signaling pathway. An intermediate must be phosphorylated on two different residues, one targeted by each of two pathways, in order to become active and produce a response. The response only occurs if the first pathway AND the second pathway are active.
Right diagram: logical "OR" in a cell signaling pathway. An intermediate must phosphorylated on a single residue in order to become active and produce a response, and either of two pathways can phosphorylate the same residue. The response occurs if the first pathway OR the second pathway is active.
Another source of complexity in signaling is that the same signaling molecule may produce different results depending on what molecules are already present in the cell. For example, the ligand acetylcholine causes opposite effects in skeletal and heart muscle because these cell types produce different kinds of acetylcholine receptors that trigger different pathways.
Cell type specificity in response to acetylcholine.
Left panel: skeletal muscle cell. The acetylcholine molecule binds to a ligand-gated ion channel, causing it to open and allowing positively charged ions to enter the cell. This event promotes muscle contraction.
Right panel: cardiac muscle cell. The acetylcholine molecule binds to a G protein-coupled receptor, triggering a downstream response that leads to inhibition of muscle contraction.
These are just a few examples of the complexities that make signaling pathways challenging, but also fascinating, to study. Cell-cell signaling pathways, especially the epidermal growth factor pathway we saw earlier, are a focus of study for researchers developing new drugs against cancer.
Want to join the conversation?
- What regulates the distribution of fluid between interstitial and intracellular compartments?(7 votes)
- I was under the impression that this is due to osmotic pressure, so differences in solute concentration(23 votes)
- What mechanism ampfily the signal ? and why they have to do that ?(5 votes)
- Many if not all of the steps described in this article can amplify a signal.
For example each step in the section titled Phosphorylation example: MAPK signaling cascade involves a kinase phosphorylating downstream molecules. For instance, if each time a RAF molecule gets activated it phosphorylates 20 molecules of MEK, then you've amplified the signal by 20 times.
Why questions are typically difficult in biology, but I'll give a slightly hypothetical example of why this is important. One response a cell needs to divide is an increased rate of protein synthesis, so maybe at the end of the pathway ERK needs to phosphorylate the (up to) 10 million ribosomes to increase their activity. This would take a very long time if only a few ERK molecules were activated! In addition, there are 244 known direct targets identified for ERK in humans§, so even if you only had a few of each of those molecules amplification would still be needed to get a timely response!
(Note: Those targets often need to modified in multiple locations — for example at least two ribosomal proteins are targets of ERK, one on two different amino acids.)
Does that help?(11 votes)
- How exactly does the cell signalling reverse?(5 votes)
- This differs per pathway. For pathways that release Ca2+ for example, the Ca2+ pumps pump the Ca2+ back into the ER, so there will be less and less Ca2+ in the cytosol.
Also, in these messenger cascades, often at the end of the cascade a phosphatase is activated, which will deactivate the kinases. If only a very short, small signal is needed, the phosphatases will be activated earlier on in the cascade (depends on the pathway).
For the ligand/receptor interactions: a ligand often binds it's receptor for a short while, but not very long, so the initial signal won't be active for long.
Hope this helps you out a bit!(8 votes)
- Do all signaling pathways simply turn on or turn off enzymes?(5 votes)
- No, they can also regulate the transcription of genes, the translation of proteins, the behavior of structural proteins, vesicle transport within cells, inhibitors of enzymes, and countless other processes. You can probably assume that most processes in a cell are affected in some way by at least one signaling pathway!(6 votes)
- can someone explain how this works with insulin and glucose? Along with GLUT2 and GLUT4 and their function in all of this? Is GLUT4 relevant for all body cells or just muscle and adipose? So confusing...(4 votes)
- GLUT4 is an insulin-responsive glucose transporter that is found in the heart, skeletal muscle, adipose tissue, and brain.
GLUT2 is expressed mainly in beta cells of the pancreas, liver, and kidney.
There are many more receptors:
Class I facilitative glucose transporters are represented by GLUT1 to GLUT4,
Class II has four members, namely, GLUT5, GLUT7, GLUT9, and GLUT11.
Class III glucose facilitative transporters, namely, GLUT6, GLUT8, GLUT10, GLUT12 and GLUT13 (HMIT).
- What does it mean to be a proto-oncogene? I have been having a hard time grasping what oncogenes are.(2 votes)
- Thank you for this question. I've recorded a video on this topic and will give you a link once I upload it.
So protooncogene is signalling molecule which acts as green light to form a tumour. It is switched on for tumour. If it is activated (or even mutated) tumour arise.
You may ask, how comes I am not having cancer?
Because proto-oncogene has its antidote - tumour suppressor gene.
Tumour suppressor gene acts as a brake for cell cycle and sits between G0 and G1 phase of the cell cycle.
No matter that you have proto-oncogenes, as long as you have an unmutated version of tumour suppressor genes, proto-oncogenes won't harm you.
You can think of them both as antigen and antibody.(2 votes)
- How does cAMP activate protein kinase A? If it's through phosphorylation, does the cAMP disappear after it has done its job (because it loses its only phosphate)?(2 votes)
- No, not through phosphorylation. Note that the text says that it "activates" PKA and the latter then establishes a phosphorylation cascade.
PKA is composed of 4 subunits - 2 Regulatory (R) and 2 Catalytic (C) subunits; and as the name suggests only the 2 C subunits have a further signaling role. The R-subunits feature cAMP-binding sites.
In the resting state when cAMP levels are low, the C-subunits are in a deactivated state due to the R-subunits. What cAMP does is that when it binds at the R sites, it causes a dissociation of the C subunits from this protein ("conformational change") making them now "activated" and thus eliciting downstream responses.
And yes the cAMP does "disappear" after it has done its job but not quite by losing its phosphate. The enzyme phosphodiesterase converts cyclic-AMP to AMP (by hydrolyzing the 3'C-Phosphate bond) marking the termination of the pathway.
- Quick question, can someone explain to me why the G Protein-Coupled receptor goes through the membrane seven times? Is this to increase the surface area in order to interact more with the membrane?(2 votes)
- It's because they have to bind lots of signals and interact on many 'fronts'.
They mediate pleiotropic transmembrane (TM) signal transduction by allosterically facilitating information transfer across the cellular membrane in response to extracellular signals [1,2] and converting them into one or more intracellular signalling cascades
Of course, by spanning many times through membrane they increase surface plus are tightly embedded into membrane.
- Is PI3 is the PI3K cell signalling? And What is TLK cell signalling?(1 vote)
- TLR stands for toll-like receptor — I found one article and one practice question mentioning this class of protiens on KhanAcademy:
Outside of KhanAcademy this free online textbook should be a good place to learn more:
These may also be useful:
- As a transcription factor...what role does c-myc play in transcription once the phosphate binds?(2 votes)