Human SPHK1 (NP_001136074.1, 1 a.a. - 384 a.a.) full-length recombinant protein with GST tag expressed in baculovirus infected Sf21 cells.
Theoretical MW (kDa):
Baculovirus infected insect cell (Sf21) expression system
Glutathione sepharose chromatography
91 % by SDS-PAGE/CBB staining
The activity was measured by off-chip mobility shift assay. The enzyme was incubated with fluorescence-labeled substrate and Mg(or Mn)/ATP. The phosphorylated and unphosphorylated substrates were separated and detected by LabChip 3000. Substrate: Sphingosine. ATP: 100 uM.
Quality Control Testing:
Loading 1 ug protein in SDS-PAGE
In 50 mM Tris-HCl, 150 mM NaCl, pH 7.5 (0.05% Brij35, 1 mM DTT, 10% glycerol)
Store at -80°C. Aliquot to avoid repeated freezing and thawing.
Sphingosine-1-phosphate (SPP) is a novel lipid messenger with both intracellular and extracellular functions. Intracellularly, it regulates proliferation and survival, and extracellularly, it is a ligand for EDG1 (MIM 601974). Various stimuli increase cellular levels of SPP by activation of sphingosine kinase (SPHK), the enzyme that catalyzes the phosphorylation of sphingosine. Competitive inhibitors of SPHK block formation of SPP and selectively inhibit cellular proliferation induced by a variety of factors, including platelet-derived growth factor (e.g., MIM 173430) and serum.[supplied by OMIM