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Related Concept Videos

SNAREs and Membrane Fusion01:43

SNAREs and Membrane Fusion

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Once a transport vesicle has recognized its target organelle, the vesicular membrane needs to fuse with the target membrane to unload the cargo. Transmembrane proteins called SNAREs present on organelle membranes and their vesicles, mediate vesicle fusion.
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Cycloaddition Reactions: MO Requirements for Thermal Activation01:16

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Thermal cycloadditions are reactions where the source of activation energy needed to initiate the reaction is provided in the form of heat. A typical example of a thermally-allowed cycloaddition is the Diels–Alder reaction, which is a [4 + 2] cycloaddition. In contrast, a [2 + 2] cycloaddition is thermally forbidden.
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Mechanisms of Membrane Domain Formation00:59

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Different physical properties of lipids and proteins allow them to localize and form distinct islands or domains in the membrane. Some membrane domains are formed due to protein-protein interactions, whereas others are formed due to the presence of specific lipids such as sphingolipids and sterols—for example, large proteins, such as bacteriorhodopsin, aggregate and create distinct domains.
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Biological membranes are more than just a barrier separating cell cytoplasm from the outside environment. They are highly dynamic and help maintain the integrity and physiological stability of the cells as well as membrane-bound organelles. Membranes also play vital roles in cell-to-cell and intracellular communication.
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Fusion of Secretory Vesicles with the Plasma Membrane01:26

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Proteins and neurotransmitters in secretory vesicles can be released from a cell upon vesicle docking, priming, and fusion with the plasma membrane. Vesicles are docked and primed in preparation for the quick exocytosis of their contents in response to a stimulus. The fusion process is mainly carried out by a SNAP Receptor or SNARE complex, consisting of synaptobrevin, syntaxin-1, and SNAP-25.
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SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy
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Strain-Promoted Cycloadditions in Lipid Bilayers Triggered by Liposome Fusion.

Coline Jumeaux1, Christopher D Spicer1,2,3, Patrick Charchar4

  • 1Department of Materials, Department of Bioengineering, and Institute of Biomedical Engineering, Imperial College London, London, SW7 2AZ, United Kingdom.

Angewandte Chemie (International Ed. in English)
|March 4, 2024
PubMed
Summary

This study explores the strain promoted azide-alkyne cycloaddition (SPAAC) reaction within liposomal membranes. Researchers developed a novel method using Förster resonance energy transfer (FRET) dyes to characterize SPAAC, enabling new bioorthogonal labeling and biosensing applications.

Keywords:
FRETSPAACbioconjugationliposomesmolecular dynamics

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Membrane Biophysics

Background:

  • Cell membranes are complex and challenging to study due to their diverse functions.
  • Bioorthogonal reactions, like strain promoted azide-alkyne cycloaddition (SPAAC), are valuable for studying biomolecules in vivo.
  • SPAAC has been underexplored in the context of lipid bilayers.

Purpose of the Study:

  • To develop and characterize a novel approach for studying SPAAC reactions within liposomal membranes.
  • To investigate the SPAAC reaction kinetics between diffusing molecules in a lipid bilayer environment.
  • To establish a foundation for in situ bioorthogonal labeling and biosensing applications in membranes.

Main Methods:

  • Utilized azide- and strained alkyne-functionalized Förster resonance energy transfer (FRET) dye pairs.
  • Developed liposomal membrane models to mimic native cellular environments.
  • Characterized the SPAAC reaction dynamics within these liposomal systems.

Main Results:

  • Successfully demonstrated and characterized the SPAAC reaction occurring between diffusing molecules inside liposomal membranes.
  • Provided the first in-depth analysis of SPAAC within a lipid bilayer context.
  • Established a new FRET-based methodology for studying bioorthogonal reactions in membranes.

Conclusions:

  • The developed FRET-based method enables the study of SPAAC reactions in liposomal membranes.
  • This research opens avenues for in situ bioorthogonal labeling of membrane proteins.
  • Potential applications include enhanced understanding of membrane dynamics and development of novel biosensors.