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Microsomal CTP:choline phosphate cytidylyltransferase: kinetic mechanism of fatty acid stimulation.
- Source :
-
Biochimica et biophysica acta [Biochim Biophys Acta] 1991 Oct 15; Vol. 1086 (1), pp. 57-62. - Publication Year :
- 1991
-
Abstract
- Fatty acids are known to cause an increase in the incorporation of radioactive choline into phosphatidylcholine. A coincident increase in membrane cytidylyltransferase activity is well documented. The purpose of the present studies was to determine the direct effects of oleic acid on the kinetic properties of membrane cytidylyltransferase. An examination of the reaction characteristics of membrane cytidylyltransferase revealed that membranes from adult rat lung contained high CTPase activity. This activity prevented the determination of reaction velocities at low CTP concentrations. The CTPase activity was blocked by the addition of ADP or ATP to the reaction. The addition of 6.0 mM ADP to the assay mixture enabled us to determine the effect of oleate on the CTP Km. Oleate (122 microM) caused a significant decrease in CTP Km for microsomal cytidylyltransferase (0.99 mM to 0.33 mM) and H-Form cytidylyltransferase (1.04 mM to 0.27 mM). Oleate did not decrease the CTP Km for L-Form cytidylyltransferase. Oleate had no effect on the choline phosphate Km in microsomal, H-Form or L-Form cytidylyltransferase. Oleate also increased the Vmax for cytidylyltransferase. The increase was dependent upon the concentration of oleate with a maximal increase of 50-60% at 100-130 microM oleate. We conclude that oleate has a direct stimulatory effect on cytidylyltransferase when it is in the active form (membrane bound or H-Form lipoprotein complex). We suggest that the kinetic effects operate synergistically with other regulatory mechanisms such as translocation or conversion of inactive to active species. The direct effect of oleate on the cytidylyltransferase may be an important regulatory mechanism when CTP concentrations are limiting.
- Subjects :
- Animals
Centrifugation, Density Gradient
Choline-Phosphate Cytidylyltransferase
In Vitro Techniques
Kinetics
Lung enzymology
Male
Microsomes, Liver enzymology
Nucleotidyltransferases metabolism
Oleic Acid
Oleic Acids pharmacology
Rats
Fatty Acids pharmacology
Microsomes enzymology
Nucleotidyltransferases drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0006-3002
- Volume :
- 1086
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Biochimica et biophysica acta
- Publication Type :
- Academic Journal
- Accession number :
- 1659454
- Full Text :
- https://doi.org/10.1016/0005-2760(91)90154-a