Characterization of the fatty acid-sensitive glucose 6-phosphate dehydrogenase from Pseudomonas cepacia.
The adenosone 5'-triphosphate-insensitive glucose 6-phosphate dehydrogenase from Pseudomonas cepacia has been found to be strongly inhibited by long-chain fatty acids and their acyl coenzyme A esters, suggesting that an important role of this isoenzyme might be to provide reduced nicotinamide adenine dinucleotide phosphate for reductive steps in fatty acid synthesis. The enzyme, which has been redesignated the fatty acid-sensitive glucose 6-phosphate dehydrogenase, has been purified to homogeneity using affinity chromatography with nicotinamide adenine dinulceotide phosphate-substituted Sepharose as a key step in the purification. The purified preparations were used to study the immunological properties and subunit composition of the enzyme and its relationship to the adenosine 5'-triphosphate-sensitive glucose 6-phosphate dehydrogenase present in extracts of P. cepacia. Although both enzymes were found to be composed of similar size subunits of about 60,000 daltons, immunological studies failed to demonstrate any antigenic similarity between them. Studies of the sedimentation behavior of the fatty acid-sensitive enzyme in sucrose gradients indicated that its apparent molecular weight is increased in the presence of glucose 6-phosphate and suggest that it may exist in an aggregated state in vivo. Palmitoyl coenzyme A, which strongly inhibited the enzyme, failed to influence its sedimentation behavior.
D000214:Acyl Coenzyme A / Q000494:pharmacology; D002846:Chromatography, Affinity; D000939:Epitopes; D005227:Fatty Acids / Q000494:pharmacology; D005954:Glucosephosphate Dehydrogenase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology*
D000214:Acyl Coenzyme A; D000939:Epitopes; D005227:Fatty Acids; D005958:Glucosephosphates; D007527:Isoenzymes; D009243:NAD; D009249:NADP; D005954:Glucosephosphate Dehydrogenase