TY - JOUR
T1 - Sphingosine inhibits nitric oxide synthase from cerebellar granule cells differentiated in vitro
AU - Viani, Paola
AU - Giussani, Paola
AU - Riboni, Laura
AU - Bassi, Rosaria
AU - Tettamanti, Guido
PY - 1999/7/9
Y1 - 1999/7/9
N2 - The effects of different bioactive sphingoid molecules on NOS activity of differentiated cerebellar granule cells were investigated by measuring the conversion of [3H]arginine to [3H]citrulline. Cytosolic Ca2+-dependent NOS activity was strongly inhibited in a dose-dependent manner by sphingosine in concentrations of 1-40 μM. This inhibition seems to be peculiar to sphingosine in that ceramide, N-acetylsphingosine, sphingosine-1P, sphinganine and tetradecylamine have no effect on the cytosolic enzyme at the considered concentrations, suggesting that it is the bulk of the sphingosine hydrophilic portion that is critical for cytosolic NOS inhibition. This inhibition of cytosolic NOS is not reversed by increasing the arginine concentration, so a competitive mechanism can be excluded. Instead, increasing the concentrations of calmodulin led to loss of sphingosine inhibition, suggesting that sphingosine interferes with the calmodulin-dependent activation of the enzyme by a competitive mechanism. Sphingosine and related compounds had no effect on the particulate Ca2+-independent NOS activity. The data obtained suggest that sphingosine could be involved in the regulation of NO production in neurons. Copyright (C) 1999 Federation of European Biochemical Societies.
AB - The effects of different bioactive sphingoid molecules on NOS activity of differentiated cerebellar granule cells were investigated by measuring the conversion of [3H]arginine to [3H]citrulline. Cytosolic Ca2+-dependent NOS activity was strongly inhibited in a dose-dependent manner by sphingosine in concentrations of 1-40 μM. This inhibition seems to be peculiar to sphingosine in that ceramide, N-acetylsphingosine, sphingosine-1P, sphinganine and tetradecylamine have no effect on the cytosolic enzyme at the considered concentrations, suggesting that it is the bulk of the sphingosine hydrophilic portion that is critical for cytosolic NOS inhibition. This inhibition of cytosolic NOS is not reversed by increasing the arginine concentration, so a competitive mechanism can be excluded. Instead, increasing the concentrations of calmodulin led to loss of sphingosine inhibition, suggesting that sphingosine interferes with the calmodulin-dependent activation of the enzyme by a competitive mechanism. Sphingosine and related compounds had no effect on the particulate Ca2+-independent NOS activity. The data obtained suggest that sphingosine could be involved in the regulation of NO production in neurons. Copyright (C) 1999 Federation of European Biochemical Societies.
KW - Ceramide
KW - Cerebellar granule cell
KW - Neuronal differentiation
KW - Nitric oxide synthase
KW - Sphingosine
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UR - http://www.scopus.com/inward/citedby.url?scp=0033008219&partnerID=8YFLogxK
U2 - 10.1016/S0014-5793(99)00836-4
DO - 10.1016/S0014-5793(99)00836-4
M3 - Article
C2 - 10431831
AN - SCOPUS:0033008219
VL - 454
SP - 321
EP - 324
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 3
ER -