Characterization of rhodosaminyl transfer by the AknS/AknT glycosylation complex and its use in reconstituting the biosynthetic pathway of aclacinomycin A

Catherine Leimkuhler, Micha Fridman, Tania Lupoli, Suzanne Walker, Christopher T. Walsh, Daniel Kahne

Research output: Contribution to journalArticle

Abstract

The tetracyclic core of anthracycline natural products with antitumor activity such as aclacinomycin A are tailored during biosynthesis by regioselective glycosylation. We report the first synthesis of TDP-L-rhodosamine and demonstrate that the glycosyltransferase AknS transfers L-rhodosamine to the aglycone to initiate construction of the side-chain trisaccharide. The partner protein AknT accelerates AknS turnover rate for L-rhodosamine transfer by 200-fold. AknT does not affect the Km but rather affects the kcat. Using these data, we propose that AknT causes a conformational change in AknS that stabilizes the transition state and ultimately enhances transfer. When the subsequent glycosyltransferase AknK and its substrate TDP-L-fucose are also added to the aglycone, the disaccharide and low levels of a fully reconstituted trisaccharide form of aclacinomycin are observed.

Original languageEnglish (US)
Pages (from-to)10546-10550
Number of pages5
JournalJournal of the American Chemical Society
Volume129
Issue number34
DOIs
StatePublished - Aug 29 2007

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Aclarubicin
Glycosylation
Trisaccharides
Glycosyltransferases
Biosynthesis
Biosynthetic Pathways
Proteins
Fucose
Disaccharides
Anthracyclines
Substrates
Biological Products

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Characterization of rhodosaminyl transfer by the AknS/AknT glycosylation complex and its use in reconstituting the biosynthetic pathway of aclacinomycin A. / Leimkuhler, Catherine; Fridman, Micha; Lupoli, Tania; Walker, Suzanne; Walsh, Christopher T.; Kahne, Daniel.

In: Journal of the American Chemical Society, Vol. 129, No. 34, 29.08.2007, p. 10546-10550.

Research output: Contribution to journalArticle

Leimkuhler, Catherine ; Fridman, Micha ; Lupoli, Tania ; Walker, Suzanne ; Walsh, Christopher T. ; Kahne, Daniel. / Characterization of rhodosaminyl transfer by the AknS/AknT glycosylation complex and its use in reconstituting the biosynthetic pathway of aclacinomycin A. In: Journal of the American Chemical Society. 2007 ; Vol. 129, No. 34. pp. 10546-10550.
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AU - Kahne, Daniel

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