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EC number: 203-710-0
CAS number: 109-83-1
1. Time-course of [3H]MME incorporation into phospholipids and
Fetal rat brain aggregating cultures were grown in medium containing
labeled MME for different time intervals and the radioactivity
incorporated into the lipids plateaued at about 48 hours.
a) The principal labeled phospholipid was
phosphatidylmonomethylethanolamine (PMME) which contained more than 80%
of the total lipid bound radioactivity. Little label was associated with
either phosphatidyldimethylethanolamine (PDME) 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-
MME). There was less radioactivity associated with MME and the presumed
2. Effect of Varying the Concentrations of [3H]DME on the Labeling of
Water- Solubles and Phospholipids.
The amount of radioactivity present in products in the presence of
varying concentrations of [3H]MME was estimated after an 8 hour
incubation. The amount of radioactivity present in PMME and Ph-MME
continued to increase up to 4 mM [3H]MME, the highest concentration
employed. The quantity of radioactivity in PDME and PC (Figure 3A) was
slight at 4 mM MME suggesting that N-methyltransferase activity is
relatively inactive under these conditions. There was a slight increase
of the labeling of MME and CDP-MME with increasing base content in the
3. Effect of Various Growth Media on MME and DME Incorporation.
a) Control = cells grown and incubated in DMEM (Dulbecco's Modified
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]MME for 24 hours, 25% of the isotope was
recovered in lipids (Table I) and most was present in PMME. In cells
grown in medium devoid of choline, the labeling of lipids and PMME 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 PMME to PDME or
PDME to PC was very low in the three growth medium. The radioactivity
associated with MME and Ph- MME 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 MME and this
resulted in a rapid decline in the radioactivity present in MME and
Ph-MME by 2 days. The decrease was less pronounced with the
phospholipids and at 2 days, PMME was still the major product.
Fetal rat brain aggregating cell cultures were exposed to varying
concentrations of [3H]monomethylethanolamine (MME) and [3H]
dimethylethanolamine (DME). 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 (PMME) and 3% in
phosphatidyldimethylethanolamine (PDME) or 95% in PDME and 5% in
phosphatidylcholine (PC) after growth in presence of [3H]MME and [3H]DME
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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