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Sterol Carrier Protein-2: Binding Protein for Endocannabinoids

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Abstract

The endocannabinoid (eCB) system, consisting of eCB ligands and the type 1 cannabinoid receptor (CB1R), subserves retrograde, activity-dependent synaptic plasticity in the brain. eCB signaling occurs “on-demand,” thus the processes regulating synthesis, mobilization and degradation of eCBs are also primary mechanisms for the regulation of CB1R activity. The eCBs, N-arachidonylethanolamine (AEA) and 2-arachidonoylglycerol (2-AG), are poorly soluble in water. We hypothesize that their aqueous solubility, and, therefore, their intracellular and transcellular distribution, are facilitated by protein binding. Using in silico docking studies, we have identified the nonspecific lipid binding protein, sterol carrier protein 2 (SCP-2), as a potential AEA binding protein. The docking studies predict that AEA and AM404 associate with SCP-2 at a putative cholesterol binding pocket with ∆G values of −3.6 and −4.6 kcal/mol, respectively. These values are considerably higher than cholesterol (−6.62 kcal/mol) but consistent with a favorable binding interaction. In support of the docking studies, SCP-2-mediated transfer of cholesterol in vitro is inhibited by micromolar concentrations of AEA; and heterologous expression of SCP-2 in HEK 293 cells increases time-related accumulation of AEA in a temperature-dependent fashion. These results suggest that SCP-2 facilitates cellular uptake of AEA. However, there is no effect of SCP-2 transfection on the cellular accumulation of AEA determined at equilibrium or the IC50 values for AEA, AM404 or 2-AG to inhibit steady state accumulation of radiolabelled AEA. We conclude that SCP-2 is a low affinity binding protein for AEA that can facilitate its cellular uptake but does not contribute significantly to intracellular sequestration of AEA.

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Abbreviations

2-AG:

2-Arachidonoylglycerol

AEA:

N-Arachidonylethanolamine

ANOVA:

Analysis of variance

CB1R:

Type 1 cannabinoid receptor

eCBs:

Endocannabinoids

ECS:

Endocannabinoid signaling

SCP-2:

Sterol carrier protein 2

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Acknowledgements

These studies were supported by NIH grants DA09155 and DA026996 and by the Advancing a Healthier Wisconsin Endowment of the Medical College of Wisconsin. CWC was supported by a New Investigator Award from the American Association of Colleges of Pharmacy (AACP).

Conflicts of Interest and Roles in the Study

None of the authors have any conflicts of interest to report. ESL carried out the biological studies; CDV, DSS and CWC carried out the docking studies. ESL, DSS, CWC and CJH conceived the studies and wrote the manuscript.

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Authors and Affiliations

  1. Neuroscience Research Center and Departments of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Elizabeth Sabens Liedhegner & Cecilia J. Hillard

  2. Department of Pharmaceutical Sciences, Concordia University of Wisconsin, School of Pharmacy, Mequon, WI, 53097, USA

    Caleb D. Vogt, Daniel S. Sem & Christopher W. Cunningham

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  1. Elizabeth Sabens Liedhegner
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  2. Caleb D. Vogt
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  3. Daniel S. Sem
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  5. Cecilia J. Hillard
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Correspondence to Cecilia J. Hillard.

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Liedhegner, E.S., Vogt, C.D., Sem, D.S. et al. Sterol Carrier Protein-2: Binding Protein for Endocannabinoids. Mol Neurobiol 50, 149–158 (2014). https://doi.org/10.1007/s12035-014-8651-7

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