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bioRxiv · 10.1101/2022.09.10.507427

Ceramide-1-phosphate transfer protein enhances lipid transport by disrupting hydrophobic lipid-membrane contacts

Rogers, J. R.·Geissler, P. L.
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Abstract

Cellular distributions of the sphingolipid ceramide-1-phosphate (C1P) impact essential biological processes. C1P levels are spatiotemporally regulated by ceramide-1-phosphate transfer protein (CPTP), which efficiently shuttles C1P between organelle membranes. Yet, how CPTP rapidly extracts and inserts C1P into a membrane remains unknown. Here, we devise a multiscale simulation approach to elucidate biophysical details of CPTP-mediated C1P transport. We find that CPTP binds a membrane poised to extract and insert C1P and that membrane binding promotes conformational changes in CPTP that facilitate C1P uptake and release. By significantly disrupting a lipids local hydrophobic environment in the membrane, CPTP lowers the activation free energy barrier for passive C1P desorption and enhances C1P extraction from the membrane. Upon uptake of C1P, further conformational changes may aid membrane unbinding in a manner reminiscent of the electrostatic switching mechanism used by other lipid transfer proteins. Insertion of C1P into an acceptor membrane, eased by a decrease in membrane order by CPTP, restarts the transfer cycle. Most notably, we provide molecular evidence for CPTPs ability to catalyze C1P extraction by breaking hydrophobic C1P-membrane contacts with compensatory hydrophobic lipid-protein contacts. Our work, thus, provides biophysical insights into how CPTP efficiently traffics C1P between membranes to maintain sphingolipid homeostasis and, additionally, presents a simulation method aptly suited for uncovering the catalytic mechanisms of other lipid transfer proteins. Author summaryCritical cellular processes require spatiotemporal regulation of sphingolipid levels among organelle membranes. Programmed cell death and inflammation, for example, are impacted by the distribution of ceramide-1-phosphate (C1P). C1P levels are specifically altered by ceramide-1-phosphate transfer protein (CPTP), which mediates C1P intermembrane transport. Using a multiscale simulation approach tailored to studying lipid transport, we elucidate key steps in the molecular mechanism used by CPTP to rapidly transport C1P between membranes: Through conformational changes that are coupled to membrane binding, CPTP significantly disrupts C1Ps local hydrophobic environment in a membrane and catalyzes its extraction. Since this catalytic mechanism is biophysically related to that of passive lipid transport, it may be ubiquitously used by lipid transport proteins to rapidly traffic lipids between membranes and ensure membrane homeostasis. Our multiscale simulation approach offers a framework to test this hypothesis and, thus, further our molecular knowledge of how lipid transfer proteins function to regulate cellular lipid distributions.

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Rogers, J. R., Geissler, P. L.. 2022-09-11. Ceramide-1-phosphate transfer protein enhances lipid transport by disrupting hydrophobic lipid-membrane contacts. https://doi.org/10.1101/2022.09.10.507427

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