Engineering of an active animal fatty acid synthase dimer with only one competent subunit.
Animal fatty acid synthases are large polypeptides containing seven functional domains that are active only in the dimeric form. Inactivity of the monomeric form has long been attributed to the obligatory participation of domains from both subunits in catalysis of substrate loading and condensation reactions. However, we have engineered a fatty acid synthase containing one wild-type subunit and one subunit compromised by mutations in all seven functional domains that is active in fatty acid synthesis. This finding indicates that a single subunit, in the context of a dimer, is able to catalyze the entire biosynthetic pathway and suggests that, in the natural complex, each of the two subunits forms a scaffold that optimizes the conformation of the companion subunit.
Joshi
Anil K
AK
Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
Rangan
Vangipuram S
VS
Witkowski
Andrzej
A
Smith
Stuart
S
D000096:Acetone / Q000031:analogs & derivatives* / Q000737:chemistry; D000818:Animals; D015153:Blotting, Western; D002384:Catalysis; D003432:Cross-Linking Reagents; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D064429:Fatty Acid Synthases / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D006358:Hot Temperature; D007202:Indicators and Reagents; D008968:Molecular Conformation; D009154:Mutation / Q000235:genetics; D015202:Protein Engineering
D003432:Cross-Linking Reagents; D018076:DNA, Complementary; D007202:Indicators and Reagents; D000096:Acetone; C047048:1,3-dibromo-2-propanone; D064429:Fatty Acid Synthases