Abstract
Acyl carrier protein (ACP) is a required cofactor for fatty acid synthesis in Escherichia coli. Mutants lacking beta-ketoacyl-ACP synthase II activity (fabF1 or fabF3) possessed a different molecular species of ACP (F-ACP) that was separated from the normal form of the protein by conformationally sensitive gel electrophoresis. Synthase I mutants contained the normal protein. Complementation of fabF1 mutants with an F' factor harboring the wild-type synthase II allele resulted in the appearance of normal ACP, whereas complementation with an F' possessing the fabF2 allele (a mutation that produces a synthase II enzyme with altered catalytic activity) resulted in the production of both forms of ACP. The structural difference between F-ACP and ACP persisted after the removal of the 4'-phosphopantetheine prosthetic group, and both forms of the protein had identical properties in an in vitro fatty acid synthase assay. Both ACP and F-ACP were purified to homogeneity, and their primary amino acid sequences were determined. The two ACP species were identical but differed from the sequence reported for E. coli E-15 ACP in that an Asn instead of an Asp was at position 24 and an Ile instead of a Val was at position 43. Therefore, F-ACP appears to be a modification of ACP that is detected when beta-ketoacyl-ACP synthase II activity is impaired.
MeSH Terms
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase/genetics,metabolism
Acyl Carrier Protein/analysis,isolation & purification,metabolism
Acyltransferases/genetics
Amino Acid Sequence
Escherichia coli/analysis,enzymology,genetics
Fatty Acids/biosynthesis
Genetic Complementation Test
Mutation
Oleic Acids/biosynthesis
Chemicals
Acyl Carrier Protein
Fatty Acids
Oleic Acids
cis-vaccenic acid
Acyltransferases
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Jackowski S
Rock C O
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28 references, click to expand
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