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EC number: 203-710-0
CAS number: 109-83-1
1. Time-course of [3H]MMEA incorporation
into phospholipids and water-soluble products.
Fetal rat brain aggregating cultures were
grown in medium containing labeled MMEA for different time intervals and
the radioactivity incorporated into the lipids plateaued at about 48
a) The principal labeled phospholipid was
phosphatidylmonomethylethanolamine (PMMEA) which contained more than 80%
of the total lipid bound radioactivity. Little label was associated with
either phosphatidyldimethylethanolamine (PDMAE) or phosphatidylcholine
b) The magnitude of radioactivity present in
the total water-solubles was considerably greater than that present in
lipids and a plateau was reached by 24 hours. The majority of the
radioactivity was present in a material chromatogramming like
phosphorylmonomethylethanolamine (Ph- MMEA). There was less
radioactivity associated with MMEA and the presumed
2. Effect of Varying the Concentrations
of [3H]DMAE on the Labeling of Water- Solubles and Phospholipids.
The amount of radioactivity present in
products in the presence of varying concentrations of [3H]MMEA was
estimated after an 8 hour incubation. The amount of radioactivity
present in PMMEA and Ph-MMEA continued to increase up to 4 mM [3H]MMEA,
the highest concentration employed. The quantity of radioactivity in
PDMAE and PC (Figure 3A) was slight at 4 mM MMEA suggesting that
N-methyltransferase activity is relatively inactive under these
conditions. There was a slight increase of the labeling of MMEA and
CDP-MMEA with increasing base content in the medium.
3. Effect of Various Growth Media on MME
and DME Incorporation.
a) Control = cells grown and incubated in
DMEM (Dulbecco's Modified Eagle Medium).
b) Met = cells grown and incubated in DMEM
in L-methionine free medium
c) Ch = cells grown and incubated in DMEM in
choline free medium.
After labeling with [3H]MMEA for 24 hours,
25% of the isotope was recovered in lipids (Table I) and most was
present in PMMEA. In cells grown in medium devoid of choline, the
labeling of lipids and PMMEA was 145% and 150% respectively as compared
to control values. There was no significant differences in labeling in
cells grown in medium devoid methionine as compared to controls. The
conversion of PMMEA to PDMAE or PDMAE to PC was very low in the three
growth medium. The radioactivity associated with MMEA and Ph- MMEA was
reduced in cells grown in the absence of methionine, after 1 day the
radioactive medium was removed and replaced by DMEM medium containing
non-radioactive MMEA and this resulted in a rapid decline in the
radioactivity present in MMEA and Ph-MMEA by 2 days. The decrease was
less pronounced with the phospholipids and at 2 days, PMMEA was still
the major product.
Fetal rat brain aggregating cell cultures
were exposed to varying concentrations of [3H]monomethylethanolamine
(MMEA) and [3H] dimethylethanolamine (DMAE). The rate of labeling of
water-soluble compounds was more rapid and the amount of radioactivity
present was greater than in the lipids. After a 72 hour incubation in
the presence of millimolar concentrations of these nitrogenous bases,
the major water-soluble products were the phosphorylated form of the
bases. Little label was associated with the free bases or their cytidyl
derivate. In the phospholipids, 97% of the radioactivity was recovered
in phosphatidylmonomethylethanolamine (PMMEA) and 3% in
phosphatidyldimethylethanolamine (PDMAE) or 95% in PDMAE and 5% in
phosphatidylcholine (PC) after growth in presence of [3H]MMEA and
[3H]DMAE, respectively. The rate of formation of the radioactive
products increased as function of the concentration of the nitrogenous
base added up to 4 mM, the highest concentration employed. There was no
significant difference in the pattern of labeling with cells grown in
media devoid of methionine or choline. The turnover of the water-soluble
metabolites was more rapid than in the phospholipids where an apparent
half-life of 24 hours was calculated.
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