Global Mapping of Protein–Lipid Interactions by Using Modified Choline‐Containing Phospholipids Metabolically Synthesized in Live Cells

The protein–lipid interaction is an essential metabolic process that mediates cellular signaling and functions. Existing strategies for large‐scale mapping studies of the protein–lipid interaction fall short in their incompatibility with metabolic incorporation or inability to remove unwanted interf...

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Published inAngewandte Chemie International Edition Vol. 56; no. 21; pp. 5829 - 5833
Main Authors Wang, Danyang, Du, Shubo, Cazenave‐Gassiot, Amaury, Ge, Jingyan, Lee, Jun‐Seok, Wenk, Markus R., Yao, Shao Q.
Format Journal Article
LanguageEnglish
Published Germany Wiley Subscription Services, Inc 15.05.2017
EditionInternational ed. in English
Subjects
Alkynes
Choline
click chemistry
Fatty acids
Incompatibility
Lipids
Mammalian cells
Metabolism
Peptide mapping
phosphatidylcholine
Phospholipids
photo-crosslinking
Proteins
protein–lipid interactions
Proteomics
quantitative proteomics
Synthesis
click chemistry
photo-crosslinking
protein-lipid interactions
quantitative proteomics
phosphatidylcholine
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Summary:The protein–lipid interaction is an essential metabolic process that mediates cellular signaling and functions. Existing strategies for large‐scale mapping studies of the protein–lipid interaction fall short in their incompatibility with metabolic incorporation or inability to remove unwanted interferences from lipidated proteins. By incorporating an alkyne‐containing choline head group and a diazirine‐modified fatty acid simultaneously into choline‐containing phospholipids synthesized from live mammalian cells, protein–phospholipid interactions have been successfully imaged in live cells. Subsequent in situ profiling of the modified Cho phospholipid‐crosslinked proteins followed by quantitative proteomics allowed identification of several hundred putative phospholipid‐interacting proteins, some of which were further validated. Double the load: A novel double incorporation strategy for metabolic biosynthesis of bifunctional choline‐containing phospholipids (for example phosphatidylcholine (PC); see picture) was developed. This strategy offers significant improvement for global mapping of genuine protein–lipid interactions from live mammalian cells.
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ISSN:1433-7851
1521-3773
1521-3773
DOI:10.1002/anie.201702509