title: Metabolic perturbation of an essential pathway: evaluation of a glycine precursor of coenzyme A.
abstract: Pantetheine and its corresponding disulfide pantethine play a key role in metabolism as building blocks of coenzyme A (CoA), an essential cofactor utilized in ~4% of primary metabolism and central to fatty acid, polyketide, and nonribosomal peptide synthases. Using a combination of recombinant engineering and chemical synthesis, we show that the disulfide of N-pantoylglycyl-2-aminoethanethiol (GlyPan), with one fewer carbon than pantetheine, can rescue a mutant E. coli strain MG1655ΔpanC lacking a functional pantothenate synthetase. Using mass spectrometry, we show that the GlyPan variant is accepted by the downstream CoA biosynthetic machinery, ultimately being incorporated into essential acyl carrier proteins. These findings point to further flexibility in CoA-dependent pathways and offer the opportunity to incorporate orthogonal analogues.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0358, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Rothmann
authors.forename: MinJin
authors.initials: M
authors.lastname: Kang
authors.forename: Reymundo
authors.initials: R
authors.lastname: Villa
authors.forename: Ioanna
authors.initials: I
authors.lastname: Ntai
authors.forename: James J
authors.initials: JJ
authors.lastname: La Clair
authors.forename: Neil L
authors.initials: NL
authors.lastname: Kelleher
authors.forename: Eli
authors.initials: E
authors.lastname: Chapman
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkart
mesh_terms: D000213:Acyl Carrier Protein / Q000378:metabolism; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000096:biosynthesis / Q000378:metabolism*; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005998:Glycine / Q000378:metabolism*; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways; D008969:Molecular Sequence Data; D010204:Pantetheine / Q000031:analogs & derivatives / Q000378:metabolism; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D014508:Urea / Q000737:chemistry
chemical_list: D000213:Acyl Carrier Protein; D004220:Disulfides; D010204:Pantetheine; C005425:pantethine; D014508:Urea; D017853:Phosphotransferases (Alcohol Group Acceptor); C023088:pantothenate kinase; D010453:Peptide Synthases; C022318:pantothenate synthetase; D003065:Coenzyme A; D005998:Glycine
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