Live-cell lipid biochemistry reveals a role of diacylglycerol side-chain composition for cellular lipid dynamics and protein affinities
Every cell produces thousands of distinct lipid species, but insight into how lipid chemical diversity contributes to biological signaling is lacking, particularly because of a scarcity of methods for quantitatively studying lipid function in living cells. Using the example of diacylglycerols, promi...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2020-04, Vol.117 (14), p.7729-7738 |
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| creator | Schuhmacher, Milena Grasskamp, Andreas T. Barahtjan, Pavel Wagner, Nicolai Lombardot, Benoit Schuhmacher, Jan S. Sala, Pia Lohmann, Annett Henry, Ian Shevchenko, Andrej Coskun, Ünal Walter, Alexander M. Nadler, André |
| description | Every cell produces thousands of distinct lipid species, but insight into how lipid chemical diversity contributes to biological signaling is lacking, particularly because of a scarcity of methods for quantitatively studying lipid function in living cells. Using the example of diacylglycerols, prominent second messengers, we here investigate whether lipid chemical diversity can provide a basis for cellular signal specification. We generated photo-caged lipid probes, which allow acute manipulation of distinct diacylglycerol species in the plasma membrane. Combining uncaging experiments with mathematical modeling, we were able to determine binding constants for diacylglycerol–protein interactions, and kinetic parameters for diacylglycerol transbilayer movement and turnover in quantitative live-cell experiments. Strikingly, we find that affinities and kinetics vary by orders of magnitude due to diacylglycerol side-chain composition. These differences are sufficient to explain differential recruitment of diacylglycerol binding proteins and, thus, differing downstream phosphorylation patterns. Our approach represents a generally applicable method for elucidating the biological function of single lipid species on subcellular scales in quantitative live-cell experiments. |
| doi_str_mv | 10.1073/pnas.1912684117 |
| format | Article |
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Using the example of diacylglycerols, prominent second messengers, we here investigate whether lipid chemical diversity can provide a basis for cellular signal specification. We generated photo-caged lipid probes, which allow acute manipulation of distinct diacylglycerol species in the plasma membrane. Combining uncaging experiments with mathematical modeling, we were able to determine binding constants for diacylglycerol–protein interactions, and kinetic parameters for diacylglycerol transbilayer movement and turnover in quantitative live-cell experiments. Strikingly, we find that affinities and kinetics vary by orders of magnitude due to diacylglycerol side-chain composition. These differences are sufficient to explain differential recruitment of diacylglycerol binding proteins and, thus, differing downstream phosphorylation patterns. Our approach represents a generally applicable method for elucidating the biological function of single lipid species on subcellular scales in quantitative live-cell experiments.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1912684117</identifier><identifier>PMID: 32213584</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Affinity ; Binding ; Biodiversity ; Biological Sciences ; Chains ; Composition ; Diglycerides ; Experiments ; Lipids ; Mathematical models ; Phosphorylation ; Physical Sciences ; Protein interaction ; Proteins ; Second messengers ; Species ; Species diversity</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2020-04, Vol.117 (14), p.7729-7738</ispartof><rights>Copyright © 2020 the Author(s). 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| subjects | Affinity Binding Biodiversity Biological Sciences Chains Composition Diglycerides Experiments Lipids Mathematical models Phosphorylation Physical Sciences Protein interaction Proteins Second messengers Species Species diversity |
| title | Live-cell lipid biochemistry reveals a role of diacylglycerol side-chain composition for cellular lipid dynamics and protein affinities |
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