Apolipoprotein M supports S1P production and conservation and mediates prolonged Akt activation via S1PR1 and S1PR3

Abstract

Sphingosine 1-phosphate (S1P) is one of the lipid mediators involved in diverse physiological functions. S1P circulates in blood and lymph bound to carrier proteins. Three S1P carrier proteins have been reported, albumin, apolipoprotein M (ApoM) and apolipoprotein A4 (ApoA4). The carrier-bound S1P exerts its functions via specific S1P receptors (S1PR1-5) on target cells. Previous studies showed several differences in physiological functions between albumin-bound S1P and ApoM-bound S1P. However, molecular mechanisms underlying the carrier-dependent differences have not been clarified. In addition, ApoA4 is a recently identified S1P carrier protein, and its functional differences from albumin and ApoM have not been addressed. Here, we compared the three carrier proteins in the processes of S1P degradation, release from S1P-producing cells and receptor activation. ApoM retained S1P more stable than albumin and ApoA4 in the cell culture medium when compared in the equimolar amounts. ApoM facilitated theS1P release from endothelial cells most efficiently. Furthermore, ApoM-bound S1P showed a tendency to induce prolonged activation of Akt via S1PR1 and S1PR3. These results suggest that the carrier-dependent functional differences of S1P are partly ascribed to the differences in the S1P stability, S1P-releasing efficiency and signaling duration.

Keywords: Apolipoprotein A4; Apolipoprotein M; LC–MS/MS; Sphingosine 1-phosphate.Abbreviations: ApoA4, Apolipoprotein A4; ApoM, Apolipoprotein M; CHO, Chinese hamster ovary; ERK, Extracellular signal-regulated kinase; LC–MS/MS, Liquid chromatography–tandem mass spectrometry; LPP, Lipid phosphate phosphatase; Mfsd2b, Multiple facilitator superfamily domain containing 2B; PBS, Phosphate-buffered saline; S1P, Sphingosine 1-phosphate; S1PR1, Sphingosine 1-phosphate receptor 1; S1PR3, Sphingosine 1-phosphate receptor 3; SphK, Sphingosine kinase; Spns2, Spinster homolog 2; TBS-T, Tris-buffed saline containing 0.1% Tween20.