TY - JOUR

T1 - Live-cell lipid biochemistry reveals a role of diacylglycerol side-chain composition for cellular lipid dynamics and protein affinities

AU - Schuhmacher, Milena

AU - Grasskamp, Andreas T

AU - Barahtjan, Pavel

AU - Wagner, Nicolai

AU - Lombardot, Benoit

AU - Schuhmacher, Jan S

AU - Sala, Pia

AU - Lohmann, Annett

AU - Henry, Ian

AU - Shevchenko, Andrej

AU - Coskun, Ünal

AU - Walter, Alexander M

AU - Nadler, André

N1 - Copyright © 2020 the Author(s). Published by PNAS.

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

KW - Adenosine Triphosphate/metabolism

KW - Biosensing Techniques

KW - Cell Membrane/metabolism

KW - Cell Survival

KW - Diglycerides/chemistry

KW - Isoenzymes/metabolism

KW - Kinetics

KW - Light

KW - Lipids/chemistry

KW - Models, Biological

KW - Protein Kinase C/metabolism

KW - Proteins/metabolism

KW - Signal Transduction

U2 - 10.1073/pnas.1912684117

DO - 10.1073/pnas.1912684117

M3 - Journal article

C2 - 32213584

VL - 117

SP - 7729

EP - 7738

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 14

ER -