Proteome-wide detection of phospholipid-protein interactions in mitochondria by photocrosslinking and click chemistry.
Photoactivatable lipid analogues are uniquely suited for the detection of lipid-protein interactions in biological membranes. Based on photocrosslinking, new methodology has been developed for the proteome-wide detection of lipid-protein interactions. Bifunctional lipid analogues containing a tag for click chemistry in addition to the photoactivatable moiety enable the enrichment of the crosslinked proteins that is required for subsequent identification by mass spectrometry. In principle the phospholipid interaction-based membrane protein proteomics approach is applicable to any biomembrane and any lipid. Here, we review the background and the development of the new methodology. Results obtained with photocrosslinking in purified mitochondrial membranes from the yeast Saccharomyces cerevisiae are summarized and future perspectives discussed.
Gubbens
Jacob
J
Membrane Enzymology and Chemical Biology & Organic Chemistry, Bijvoet Institute and Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. a.i.p.m.dekroon@uu.nl
de Kroon
Anton I P M
AI
D008928:Mitochondria / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D010777:Photochemistry; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D020543:Proteome
D010743:Phospholipids; D011506:Proteins; D020543:Proteome