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    Home > Biochemistry News > Biotechnology News > Sphingosine Kinase: Assay and Product Analysis

    Sphingosine Kinase: Assay and Product Analysis

    • Last Update: 2020-11-28
    • Source: Internet
    • Author: User
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    The phosphorylation of long chain sphingoid bases on the primary alcohol group occurs in cells by the action of sphingosine kinase (
    1

    3
    ). Sphingosine kinase is present in the cytosolic fraction of most cells (
    4

    7
    ) and in the membrane fraction of certain tissues and organisms (
    8
    ,
    9
    ). The reaction product, sphingosine-1-phosphate (SPP), was considered for more than 20 yr to be merely an intermediate in the catabolism of long-chain sphingoid bases to palmitaldehyde and phosphoethanolamine (
    3
    ,
    10
    ). Studies in our lab stimulated new interest in the potential roles of SPP as a second messenger. Initially, we found that exogenous SPP initiated cell division of quiescent Swiss 3T3 fibroblasts (
    11
    ) and induced inositol trisphosphate-independent release of calcium from intracellular stores (
    11
    ,
    12
    ). SPP also has been shown to affect several signal transduction pathways including phospholipase D activation (
    13
    ), stimulation of the Raf/MEK/ERK signaling pathway (
    14
    ,
    15
    ), and inhibition of ceramide-induced activation of stress-activated protein kinase (SAPK/JNK) that leads to apoptotic responses (
    15
    ). Additional effects of SPP include stimulation of tyrosine phosphorylation of focal adhesion kinase (FAK) and the cytoskeleton-associated protein paxillin (
    16
    ). These tyrosine phosphorylations are mediated through the activation of the small G protein rho, which also mediates stress fiber formation induced by SPP (
    16
    ).
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