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Lipids as Anchors

Lipids as Anchors

In the plasma membrane, the lipids forming the bilayer can also act as an anchor to tether proteins to the membrane. The three main types of lipid anchors found in eukaryotes are – prenyl groups, fatty acyl groups, and glycosylphosphatidylinositol or GPI groups. Prenyl and fatty acyl groups act as anchors on the cytosolic surface of the membrane, whereas GPI anchors proteins on the extracellular side.
The carboxy-terminal of most of the prenylated proteins, such as Ras proteins, contains...

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Bifunctional Lipid-Protein Cross-linking Efficiency and Reaction Products.

Carla Kirschbaum^1,2, H Mathilda Lennartz³, Katelyn C Cook³

¹Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford OX1 3QU, United Kingdom.

Journal of the American Chemical Society

|April 10, 2026

Summary

Bifunctional diazirine lipids show low photo-cross-linking efficiency, with only 5% forming protein-lipid links. Most probes undergo intramolecular reactions, highlighting the need for improved experimental methods in lipid biology.

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Area of Science:

Biochemistry
Chemical Biology
Molecular Biology

Background:

Bifunctional diazirine lipids are key tools for studying protein-lipid interactions via photo-cross-linking.
The actual cross-linking efficiency of these lipid probes remains largely unevaluated.
Understanding this efficiency is crucial for accurate cellular localization and interaction mapping.

Purpose of the Study:

To systematically evaluate the photo-cross-linking efficiency of bifunctional diazirine lipid probes.
To determine the upper limit of cross-linking efficiency using a well-defined protein system.
To characterize the reaction products and identify competing reaction pathways.

Main Methods:

Utilized the lipid transfer protein STARD10, which binds phospholipids with 1:1 stoichiometry.
Employed photo-activation of bifunctional diazirine lipids in complex with STARD10.
Analyzed reaction products using native and denaturing mass spectrometry.

Main Results:

Approximately 5% of photoactivated bifunctional lipids successfully formed covalent protein-lipid cross-links.
The majority of activated lipids underwent intramolecular reactions.
Intramolecular reactions yielded products with alkene, ketone, and hydroxyl functional groups.

Conclusions:

The photo-cross-linking efficiency of bifunctional diazirine lipids is limited, with significant intramolecular side reactions.
These findings provide critical context for interpreting data from lipid-protein interaction studies.
Further optimization of experimental workflows and probe design is necessary for advancing lipid biology research.