Expression in Escherichia coli, purification and characterization of two mammalian thioesterases involved in fatty acid synthesis.
Thioesterase I, a constituent domain of the multifunctional fatty acid synthase, and thioesterase II, an independent monofunctional protein, catalyse the chain-terminating reaction in fatty acid synthesis de novo at long and medium chain lengths respectively. The enzymes have been cloned and expressed in Escherichia coli under the control of the temperature-sensitive lambda repressor. The recombinant proteins are full-length catalytically competent thioesterases with specificities indistinguishable from those of the natural enzymes.
Naggert
J
J
Children's Hospital Oakland Research Institute, CA 94609.
Witkowski
A
A
Wessa
B
B
Smith
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D064429:Fatty Acid Synthases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis; D007700:Kinetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010170:Palmitoyl-CoA Hydrolase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D005227:Fatty Acids; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D064429:Fatty Acid Synthases; D013869:Thiolester Hydrolases; C022738:thioesterase II; D010170:Palmitoyl-CoA Hydrolase