Relationship between lipid flipping and EV

Extracellular vesicles

“Lipid flipping” refers to the movement of lipid molecules from one leaflet (half) of a biological membrane to the other leaflet. This process, also known as “flip-flop” or interleaflet movement, is typically slow and energetically unfavorable because the polar head groups of lipids need to pass through the hydrophobic core of the membrane. However, it is crucial for proper cellular membrane function and maintenance. Several proteins, called flippases, floppases, and scramblases, facilitate lipid flipping and maintain the asymmetric distribution of lipids between the two leaflets of the bilayer.

What are polar head groups?

“Polar head groups” refer to a part of lipid molecules. Lipid molecules generally consist of two main parts: a polar (water-soluble) head group and a nonpolar (water-insoluble) tail.

The polar head group is the part of the lipid that interacts with the aqueous environment, leading lipids to naturally form bilayers. In this structure, the polar head groups are in contact with water, while the nonpolar tails are oriented towards the interior, avoiding contact with water. This is the basic structure of biological membranes.

What is lipid asymmetry?

“Lipid asymmetry” refers to the state where lipid (lipid) molecules are not evenly distributed across both sides (leaflets) of a biological cell membrane.

The cell membrane consists of a phospholipid bilayer, and its inner and outer leaflets have different lipid compositions. This allows the cell membrane to fulfill specific functions. For example, the inner leaflet of the cell contains a higher concentration of negatively charged lipids, which can attract positively charged proteins inside the cell.

This asymmetry is maintained by specific proteins called flippases, floppases, and scramblases. These proteins facilitate the “flip-flop” of lipids and assist in the movement of lipids between the leaflets.

What is the relationship between flip-flop and extracellular vesicles (EVs)?

Extracellular vesicles (EVs) are small membrane-bound vesicles secreted by cells that play important roles in intercellular communication. Lipid flip-flop is involved in the formation and release of these EVs.

Specifically, during the process of apoptosis (cell self-destruction), certain lipids (e.g., phosphatidylserine) flip-flop from the inner side to the outer side of the cell membrane. This signals to the surrounding environment that the cell is undergoing apoptosis. Some of these lipids also appear on the surface of extracellular vesicles formed during the process of apoptosis.

The generation and release of EVs also rely on the curvature of membranes and the asymmetry of cell membranes. Lipid flip-flop helps maintain the lipid asymmetry of the cell membrane, which supports the formation of EVs. Therefore, lipid flip-flop and the formation of EVs are closely related.

Why is lipid asymmetry important?

Lipid asymmetry in the cell membrane is important for supporting the following significant biological functions:

  1. Membrane physical properties: The asymmetric distribution of lipids determines the physical properties of the cell membrane, such as fluidity, tension, curvature, etc. These properties are essential for the proper functioning of the cell membrane.
  2. Cell signaling: Lipid asymmetry plays a crucial role in intracellular and intercellular signal transduction. For example, during apoptosis (programmed cell death), the movement of the lipid phosphatidylserine from the inner to the outer side of the cell membrane serves as a signal to other cells that the cell is dying.
  3. Cell shape and movement: Lipid asymmetry is important for maintaining cell shape and enabling cell movement.
  4. Transport and uptake: Specific nutrients and signaling molecules are taken up into the cell through specific lipids. Lipid asymmetry regulates this process.

Due to these reasons, the lipid asymmetry of the cell membrane is crucial for maintaining biological functions.

Does flipping of phosphatidylserine always occur when EVs are released?

Flipping (movement from the inner to the outer side) of phosphatidylserine (PS) is commonly observed in specific situations, especially during apoptosis (programmed cell death). However, it does not occur in all extracellular vesicle (EV) releases.

The formation and release of EVs depend heavily on the cell type, state, and the surrounding environment. Some EVs, particularly those associated with apoptosis, are labeled by the exposure of PS on the outer side of the cell membrane. However, many other EVs, including a type called exosomes, do not require such PS exposure.

Therefore, flipping of phosphatidylserine is often associated with EV release, but it does not occur in all EV releases.

Is there a relationship between exosome release and lipid flipping?

Exosomes are a specific type of extracellular vesicles (EVs) that are released from a structure called multivesicular bodies (MVB) formed inside cells. MVBs derive from endosomes, and they contain small vesicles that become exosomes. When MVBs fuse with the cell membrane, these small vesicles are released as exosomes into the extracellular space.

Lipid flipping refers to the phenomenon where specific lipids move from one layer of the cell membrane to the other. This leads to lipid asymmetry, which affects the physical properties of the cell membrane and cellular signal transduction.

The direct involvement of lipid flipping in exosome release is not fully understood yet. However, lipid asymmetry may indirectly influence the fusion of MVBs with the cell membrane and the release of exosomes by controlling membrane curvature. Additionally, the signaling generated by specific lipids flipping may regulate the release of exosomes.

What signals are involved in flipping and exosome release?

The specific signals involved in lipid flipping and exosome release have not been fully elucidated, but several possibilities have been proposed.

  1. Phosphatidylserine (PS) flipping: PS is usually located in the inner leaflet of the cell membrane but can move to the outer leaflet in specific situations, such as apoptosis or activation. This can function as an “eat-me” signal and potentially induce exosome release.
  2. Protein-mediated signaling: Certain proteins (e.g., flippases and floppases) regulate lipid flip-flop and may influence the generation and release of exosomes.
  3. Activation of signaling pathways: Various biological stimuli, including cellular stress, inflammation, and cell activation, activate intracellular signaling pathways. These signals can regulate the redistribution of lipids (including flipping) and potentially promote the generation and release of exosomes. Specific molecules such as Rho GTPases, combinatorial proteins, and the Rab family regulate exosome release.
  4. Calcium signaling: Calcium ions regulate many cellular processes, and changes in their levels may modulate lipid flipping and exosome release.
  5. These signals are believed to indirectly contribute to exosome release by affecting the physical properties of the cell membrane, the physiological state of the cell, and the intra- and extracellular environments. However, the detailed mechanisms of these processes are still not fully understood, and research is ongoing.