Biochemical and Structural Insights of the Early Glycosylation Steps in Calicheamicin Biosynthesis

Changsheng Zhang; Eduard Bitto; Randal D. Goff; Shanteri Singh; Craig A. Bingman; Byron R. Griffith; Christoph Albermann; George N. Phillips; Jon S. Thorson

doi:10.1016/j.chembiol.2008.06.011

Biochemical and Structural Insights of the Early Glycosylation Steps in Calicheamicin Biosynthesis

Changsheng Zhang, Eduard Bitto, Randal D. Goff, Shanteri Singh, Craig A. Bingman, Byron R. Griffith, Christoph Albermann, George N. Phillips, Jon S. Thorson

Research output: Contribution to journal › Article › peer-review

45 Scopus citations

Abstract

The enediyne antibiotic calicheamicin (CLM) γ₁^I is a prominent antitumor agent that is targeted to DNA by a novel aryltetrasaccharide comprised of an aromatic unit and four unusual carbohydrates. Herein we report the heterologous expression and the biochemical characterization of the two "internal" glycosyltransferases CalG3 and CalG2 and the structural elucidation of an enediyne glycosyltransferase (CalG3). In conjunction with the previous characterization of the "external" CLM GTs CalG1 and CalG4, this study completes the functional assignment of all four CLM GTs, extends the utility of enediyne GT-catalyzed reaction reversibility, and presents conclusive evidence of a sequential glycosylation pathway in CLM biosynthesis. This work also reveals the common GT-B structural fold can now be extended to include enediyne GTs.

Original language	English
Pages (from-to)	842-853
Number of pages	12
Journal	Chemistry and Biology
Volume	15
Issue number	8
DOIs	https://doi.org/10.1016/j.chembiol.2008.06.011
State	Published - Aug 25 2008

Bibliographical note

Funding Information:
We thank the University of Wisconsin–Madison School of Pharmacy Analytical Facility for analytical support, Philip R. Hamann (Wyeth Research) for providing CLM α 3 I , and Gavin J. Williams for helpful discussion. B.R.G. is a postdoctoral fellow of the American Cancer Society (PF-05-016-01-CDD) and J.S.T is a University of Wisconsin H.I. Romnes Fellow. This research was supported in part by National Institutes of Health Grant CA84374 and National Cooperative Drug Discovery Group Grant U19 CA113297 from the National Cancer Institute. The General Medicine and Cancer Institute Collaborative Access Team (GM/CA-CAT) has been funded in whole or in part with federal funds from the National Cancer Institute (Y1-CO-1020) and the National Institute of General Medical Science (Y1-GM-1104). Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science, under contract W-31-109-ENG-38. The authors report competing interests. J.S.T. is a cofounder of Centrose (Madison, WI).

Keywords

CHEMBIO

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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10.1016/j.chembiol.2008.06.011

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title = "Biochemical and Structural Insights of the Early Glycosylation Steps in Calicheamicin Biosynthesis",

abstract = "The enediyne antibiotic calicheamicin (CLM) γ1I is a prominent antitumor agent that is targeted to DNA by a novel aryltetrasaccharide comprised of an aromatic unit and four unusual carbohydrates. Herein we report the heterologous expression and the biochemical characterization of the two {"}internal{"} glycosyltransferases CalG3 and CalG2 and the structural elucidation of an enediyne glycosyltransferase (CalG3). In conjunction with the previous characterization of the {"}external{"} CLM GTs CalG1 and CalG4, this study completes the functional assignment of all four CLM GTs, extends the utility of enediyne GT-catalyzed reaction reversibility, and presents conclusive evidence of a sequential glycosylation pathway in CLM biosynthesis. This work also reveals the common GT-B structural fold can now be extended to include enediyne GTs.",

keywords = "CHEMBIO",

author = "Changsheng Zhang and Eduard Bitto and Goff, {Randal D.} and Shanteri Singh and Bingman, {Craig A.} and Griffith, {Byron R.} and Christoph Albermann and Phillips, {George N.} and Thorson, {Jon S.}",

note = "Funding Information: We thank the University of Wisconsin–Madison School of Pharmacy Analytical Facility for analytical support, Philip R. Hamann (Wyeth Research) for providing CLM α 3 I , and Gavin J. Williams for helpful discussion. B.R.G. is a postdoctoral fellow of the American Cancer Society (PF-05-016-01-CDD) and J.S.T is a University of Wisconsin H.I. Romnes Fellow. This research was supported in part by National Institutes of Health Grant CA84374 and National Cooperative Drug Discovery Group Grant U19 CA113297 from the National Cancer Institute. The General Medicine and Cancer Institute Collaborative Access Team (GM/CA-CAT) has been funded in whole or in part with federal funds from the National Cancer Institute (Y1-CO-1020) and the National Institute of General Medical Science (Y1-GM-1104). Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science, under contract W-31-109-ENG-38. The authors report competing interests. J.S.T. is a cofounder of Centrose (Madison, WI). ",

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AU - Zhang, Changsheng

AU - Bitto, Eduard

AU - Goff, Randal D.

AU - Singh, Shanteri

AU - Bingman, Craig A.

AU - Griffith, Byron R.

AU - Albermann, Christoph

AU - Phillips, George N.

AU - Thorson, Jon S.

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PY - 2008/8/25

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N2 - The enediyne antibiotic calicheamicin (CLM) γ1I is a prominent antitumor agent that is targeted to DNA by a novel aryltetrasaccharide comprised of an aromatic unit and four unusual carbohydrates. Herein we report the heterologous expression and the biochemical characterization of the two "internal" glycosyltransferases CalG3 and CalG2 and the structural elucidation of an enediyne glycosyltransferase (CalG3). In conjunction with the previous characterization of the "external" CLM GTs CalG1 and CalG4, this study completes the functional assignment of all four CLM GTs, extends the utility of enediyne GT-catalyzed reaction reversibility, and presents conclusive evidence of a sequential glycosylation pathway in CLM biosynthesis. This work also reveals the common GT-B structural fold can now be extended to include enediyne GTs.

AB - The enediyne antibiotic calicheamicin (CLM) γ1I is a prominent antitumor agent that is targeted to DNA by a novel aryltetrasaccharide comprised of an aromatic unit and four unusual carbohydrates. Herein we report the heterologous expression and the biochemical characterization of the two "internal" glycosyltransferases CalG3 and CalG2 and the structural elucidation of an enediyne glycosyltransferase (CalG3). In conjunction with the previous characterization of the "external" CLM GTs CalG1 and CalG4, this study completes the functional assignment of all four CLM GTs, extends the utility of enediyne GT-catalyzed reaction reversibility, and presents conclusive evidence of a sequential glycosylation pathway in CLM biosynthesis. This work also reveals the common GT-B structural fold can now be extended to include enediyne GTs.

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ER -

Biochemical and Structural Insights of the Early Glycosylation Steps in Calicheamicin Biosynthesis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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