REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Monoclonal Anti-FLAG M2 antibody	Sigma	Cat# F3165-.2MG
Anti-β-actin antibody	Cell Signaling Technology	Cat# 3700S
Bacterial and virus strains
DH10Bac Competent Cells	Thermo Fisher Scientific	Cat# 10361012
Stellar Competent Cells	Takara	Cat# 636763
Chemicals, peptides, and recombinant proteins
n-Dodecyl-b-D-maltopyranoside (DDM)	Anatrace	Cat# D310
18:1 PI	Avanti Polar Lipids	Cat# 850149P-500μg
18:1 PI(4)P	Avanti Polar Lipids	Cat# 850151P-500μg
18:1 PI(4,5)P2	Avanti Polar Lipids	Cat# 850155P-500μg
18:1 PI(3,4,5)P3	Avanti Polar Lipids	Cat# 850156P-500μg
16:0-18:1 PA	Avanti Polar Lipids	Cat# 840857P-500mg
16:0-18:1 PS	Avanti Polar Lipids	Cat# 840034P-500mg
16:0-18:1 PG	Avanti Polar Lipids	Cat# 840457P-500mg
16:0-18:1 PE	Avanti Polar Lipids	Cat# 850757P-500mg
16:0-18:1 PC	Avanti Polar Lipids	Cat# 850457P-500mg
Sphingosine 1-phosphate	Sigma	Cat# 73914-1MG
Deuterium oxide	Sigma	Cat# 151882-100G
Lipofectamine 2000	Thermo Fisher Scientific	Cat# 11668019
ISA-2011B	MedChemExpress	Cat# HY-16937-5mg
UNC3230	Cayman Chemical	Cat# 16401-5 mg-CAY
Critical commercial assays
Human S1P ELISA kit	MyBioSource	Cat# MBS2516132
PI(4,5)P2 ELISA kit	Echelon Biosciences	Cat# K-4500
Experimental models: Cell lines
Human: 293T cells	ATCC	Cat# CRL-3216
Human: Expi293F cells	Thermo Fisher Scientific	Cat# A14527
Insect: Sf9 cells	Thermo Fisher Scientific	Cat# 11496015
Deposited data
Raw data	This paper	https://doi.org/10.17632/vcwttwj4z7.1
Recombinant DNA
pcDNA5/TO empty vector	Thermo Fisher Scientific	Cat# V103320
pHTBV1.1 with C-terminal twin-Strep and 10×His tag	This paper	N/A
pHTBV1.1-SPNS2-GFP	This paper	N/A
pHTBV1.1-truncated SPNS2	This paper	N/A
pcDNA5/TO-SPNS2-FLAG	This paper	N/A
pcDNA5/TO-truncated SPNS2-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R23-28A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R28A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R43A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R164A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R313A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(K391A)-FLAG	This paper	N/A
pcDNA5/TO-SPNS2(R532A)-FLAG	This paper	N/A
pcDNA5/TO-EGFP-SPNS2	This paper	N/A
pcDNA5/TO-EGFP-truncated SPNS2	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R23-28A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R28A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R43A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R164A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R313A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(K391A)	This paper	N/A
pcDNA5/TO-EGFP-SPNS2(R532A)	This paper	N/A
Software and algorithms
GraphPad Prism 9	GraphPad	https://www.graphpad.com/
Volocity 6.3.1	PerkinElmer	http://www.cellularimaging.com
Xcalibur 4.4	Thermo Fisher Scientific	https://thermo.flexnetoperations.com/
Proteome Discoverer 2.5	Thermo Fisher Scientific	https://thermo.flexnetoperations.com/
Lipid Search 4.2	Thermo Fisher Scientific	https://thermo.flexnetoperations.com/
MassLynx V4.1	Waters	N/A
ProteinLynx Global Server 2.5.1	Waters	N/A
DynamX 3.0	Waters	N/A
Other
Superdex 200 Increase 10/300 GL column	GE Healthcare	Cat# 28-9909-44
Acclaim PepMap 100, C18, 75 μm × 15 cm	Thermo Fisher Scientific	Cat# 164568
Enzymate BEH Pepsin Column 2.1 × 30 mm	Waters	Cat# 186007233
ACQUITY UPLC BEH C18 1.7 μm 1.0x100 mm	Waters	Cat# 186002346