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A spectrophotometric method for the assay of cytidine 5'-diphospho-1,2-diacyl-sn-glycerol-dependent enzymes of phospholipid metabolism. 1978

G M Carman, and W Dowhan

Cytidine 5'-diphospho-1,2-diacyl-sn-glycerol (CDP-diglyceride) hydrolase, CDP-diglyceride:L-serine O-phosphatidyltransferase, and CDP-diglyceride:sn-glycero-3-phosphate phosphatidyltransferase all release CMP from their liponucleotide substrate, CDP-diglyceride. We have developed a spectrophotometric assay for these enzymes using CMP kinase, pyruvate kinase, and lactate dehydrogenase to couple the release of CMP with the oxidation of NADH. The assay for each of the phospholipid-dependent enzymes was found to be linear both with time and with enzyme concentration. The assay should prove useful for continuous monitoring of enzymatic activity, determination of initial rates of reaction, and detailed kinetic analysis of these enzymes. Since several enzymes and substrates are used in the coupled assay system, the method is limited to analysis of partially purified preparations lacking competing activities.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D009702 Nucleoside Diphosphate Sugars Diphosphate Sugars, Nucleoside,Sugars, Nucleoside Diphosphate
D010717 CDPdiacylglycerol-Serine O-Phosphatidyltransferase An enzyme that catalyzes the formation of phosphatidylserine and CMP from CDPdiglyceride plus serine. EC 2.7.8.8. CDP Diacylglycerol-Serine O-Phosphatidyltransferase,CDP Diglyceride Serine O-Phosphatidyltransferase,CDP-DG Synthase,Phosphatidylserine Synthase,Phosphatidylserine Synthetase,CDP DG Synthase,CDP Diacylglycerol Serine O Phosphatidyltransferase,CDP Diglyceride Serine O Phosphatidyltransferase,CDPdiacylglycerol Serine O Phosphatidyltransferase,Diacylglycerol-Serine O-Phosphatidyltransferase, CDP,O-Phosphatidyltransferase, CDP Diacylglycerol-Serine,O-Phosphatidyltransferase, CDPdiacylglycerol-Serine,Synthase, CDP-DG,Synthase, Phosphatidylserine,Synthetase, Phosphatidylserine
D010770 Phosphotransferases A rather large group of enzymes comprising not only those transferring phosphate but also diphosphate, nucleotidyl residues, and others. These have also been subdivided according to the acceptor group. (From Enzyme Nomenclature, 1992) EC 2.7. Kinases,Phosphotransferase,Phosphotransferases, ATP,Transphosphorylase,Transphosphorylases,Kinase,ATP Phosphotransferases
D011755 Pyrophosphatases A group of enzymes within the class EC 3.6.1.- that catalyze the hydrolysis of diphosphate bonds, chiefly in nucleoside di- and triphosphates. They may liberate either a mono- or diphosphate. EC 3.6.1.-. Pyrophosphatase
D003567 Cytidine Diphosphate Diglycerides The ester of diacylglycerol with the terminal phosphate of cytidine diphosphate. It serves as an intermediate in the biosynthesis of phosphatidylethanolamine and phosphatidylserine in bacteria. CDP Diacylglycerols,CDP Diglycerides,Cytidine Diphosphodiacylglycerols,CDP-Diacylglycerol,CDP Diacylglycerol,Diacylglycerols, CDP,Diglycerides, CDP,Diglycerides, Cytidine Diphosphate,Diphosphate Diglycerides, Cytidine,Diphosphodiacylglycerols, Cytidine
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005994 Glycerophosphates Any salt or ester of glycerophosphoric acid. Glycerolphosphate,Glycerophosphate,Calcium Glycerophosphate,Glycerolphosphates,Glycerophosphate, Calcium
D001407 Bacillus A genus of BACILLACEAE that are spore-forming, rod-shaped cells. Most species are saprophytic soil forms with only a few species being pathogenic. Bacillus bacterium
D013056 Spectrophotometry, Ultraviolet Determination of the spectra of ultraviolet absorption by specific molecules in gases or liquids, for example Cl2, SO2, NO2, CS2, ozone, mercury vapor, and various unsaturated compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Ultraviolet Spectrophotometry

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