Purification and properties of the fatty acids synthetase complex from Neurospora crassa, and the nature of the fas-mutation.
A procedure is described for the purification of the fatty acid synthetase complex (FAS) from Neurospora crassa. The enzyme complex has a molecular weight of 2.3 times 10(6), contains 6 mol of 4'-phosphopantetheine per mol, and on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate gives a single band, or a closely spaced doublet, which comigrates with standard myosin (molecular weight, 2 times 10(5)). Since the slightly retarded component in the doublet accounts for all protein-bound 4'-phosphopantetheine, the complex appears to be made up of 11 to 12 equally sized subunits, 6 of which carry the acyl carrier protein function. In this unusual arrangement, notably the lack of the low-molecular-weight acyl carrier protein component seen in other FAS systems, as well as in its enzymatic properties, the Neurospora FAS complex is quite similar to the yeast enzyme. The FAS complex of a saturated fatty acid-requiring mutant, previously disignated cel-, contains less than 2% of the 4'-phosphopantetheine prosthetic groups found in the wild-type complex. The leaky phenotype of this mutant, here designated fas-, is accounted for by a residual fatty acid synthesizing activity in its FAS complex, which is several-fold higher than expected from its residual content of 4'-phosphopanthetheine.
D000102:Acetyl-CoA C-Acyltransferase / Q000378:metabolism; D000217:Acyltransferases / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D064429:Fatty Acid Synthases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis; D008314:Malonates; D008970:Molecular Weight; D009154:Mutation; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology* / Q000378:metabolism; D010204:Pantetheine / Q000032:analysis; D010205:Pantothenic Acid / Q000378:metabolism; D011479:Protamines
D005227:Fatty Acids; D008314:Malonates; D011479:Protamines; D010205:Pantothenic Acid; D010204:Pantetheine; D000429:Alcohol Oxidoreductases; D000217:Acyltransferases; D000102:Acetyl-CoA C-Acyltransferase; D064429:Fatty Acid Synthases