Structure-based engineering of E. coli galactokinase as a first step toward in vivo glycorandomization

Jie Yang; Xun Fu; Jianchun Liao; Lesley Liu; Jon S. Thorson

doi:10.1016/j.chembiol.2005.04.009

Structure-based engineering of E. coli galactokinase as a first step toward in vivo glycorandomization

Jie Yang
, Xun Fu
, Jianchun Liao
, Lesley Liu
, Jon S. Thorson

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

66 Scopus citations

Abstract

In vitro glycorandomization is a rapid chemoenzymatic strategy to diversify complex natural product scaffolds. The glycorandomization sugar activation pathway is dependent upon the efficient construction of diverse sugar-1-phosphate libraries. In the context of the previously evolved GalK Y371H "gatekeeper" mutation, the active site M173L mutation described herein presents a kinase with remarkably broadened substrate range to include 28 diverse natural and unnatural sugars. Among these new substrates, 6-azido-6-deoxy-galactose and 6-azido-6-deoxy-glucose present unique chemical probes to assess the utility of an E. coli Y371H/M173L-GalK-overproducing strain to generate unnatural sugar-1-phosphates in vivo. Remarkably, the in vivo conversion of both unnatural sugars rival that demonstrated in vitro. This notable in vivo success stands as the first step toward constructing short sugar-activation pathways in vivo and, ultimately, in vivo natural-product glycorandomization.

Original language	English
Pages (from-to)	657-664
Number of pages	8
Journal	Chemistry and Biology
Volume	12
Issue number	6
DOIs	https://doi.org/10.1016/j.chembiol.2005.04.009
State	Published - 2005

Bibliographical note

Funding Information:
This contribution was supported in part by the National Institutes of Health grants AI52218, CA84374, and GM70637. We are grateful to the University of Wisconsin-Madison School of Pharmacy Analytical Facility for analytical support.

Funding

This contribution was supported in part by the National Institutes of Health grants AI52218, CA84374, and GM70637. We are grateful to the University of Wisconsin-Madison School of Pharmacy Analytical Facility for analytical support.

Funders	Funder number
National Institutes of Health (NIH)	GM70637, AI52218
National Childhood Cancer Registry – National Cancer Institute	R01CA084374

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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10.1016/j.chembiol.2005.04.009

Cite this

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title = "Structure-based engineering of E. coli galactokinase as a first step toward in vivo glycorandomization",

abstract = "In vitro glycorandomization is a rapid chemoenzymatic strategy to diversify complex natural product scaffolds. The glycorandomization sugar activation pathway is dependent upon the efficient construction of diverse sugar-1-phosphate libraries. In the context of the previously evolved GalK Y371H {"}gatekeeper{"} mutation, the active site M173L mutation described herein presents a kinase with remarkably broadened substrate range to include 28 diverse natural and unnatural sugars. Among these new substrates, 6-azido-6-deoxy-galactose and 6-azido-6-deoxy-glucose present unique chemical probes to assess the utility of an E. coli Y371H/M173L-GalK-overproducing strain to generate unnatural sugar-1-phosphates in vivo. Remarkably, the in vivo conversion of both unnatural sugars rival that demonstrated in vitro. This notable in vivo success stands as the first step toward constructing short sugar-activation pathways in vivo and, ultimately, in vivo natural-product glycorandomization.",

author = "Jie Yang and Xun Fu and Jianchun Liao and Lesley Liu and Thorson, \{Jon S.\}",

note = "Funding Information: This contribution was supported in part by the National Institutes of Health grants AI52218, CA84374, and GM70637. We are grateful to the University of Wisconsin-Madison School of Pharmacy Analytical Facility for analytical support.",

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doi = "10.1016/j.chembiol.2005.04.009",

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AU - Yang, Jie

AU - Fu, Xun

AU - Liao, Jianchun

AU - Liu, Lesley

AU - Thorson, Jon S.

N1 - Funding Information: This contribution was supported in part by the National Institutes of Health grants AI52218, CA84374, and GM70637. We are grateful to the University of Wisconsin-Madison School of Pharmacy Analytical Facility for analytical support.

PY - 2005

Y1 - 2005

N2 - In vitro glycorandomization is a rapid chemoenzymatic strategy to diversify complex natural product scaffolds. The glycorandomization sugar activation pathway is dependent upon the efficient construction of diverse sugar-1-phosphate libraries. In the context of the previously evolved GalK Y371H "gatekeeper" mutation, the active site M173L mutation described herein presents a kinase with remarkably broadened substrate range to include 28 diverse natural and unnatural sugars. Among these new substrates, 6-azido-6-deoxy-galactose and 6-azido-6-deoxy-glucose present unique chemical probes to assess the utility of an E. coli Y371H/M173L-GalK-overproducing strain to generate unnatural sugar-1-phosphates in vivo. Remarkably, the in vivo conversion of both unnatural sugars rival that demonstrated in vitro. This notable in vivo success stands as the first step toward constructing short sugar-activation pathways in vivo and, ultimately, in vivo natural-product glycorandomization.

AB - In vitro glycorandomization is a rapid chemoenzymatic strategy to diversify complex natural product scaffolds. The glycorandomization sugar activation pathway is dependent upon the efficient construction of diverse sugar-1-phosphate libraries. In the context of the previously evolved GalK Y371H "gatekeeper" mutation, the active site M173L mutation described herein presents a kinase with remarkably broadened substrate range to include 28 diverse natural and unnatural sugars. Among these new substrates, 6-azido-6-deoxy-galactose and 6-azido-6-deoxy-glucose present unique chemical probes to assess the utility of an E. coli Y371H/M173L-GalK-overproducing strain to generate unnatural sugar-1-phosphates in vivo. Remarkably, the in vivo conversion of both unnatural sugars rival that demonstrated in vitro. This notable in vivo success stands as the first step toward constructing short sugar-activation pathways in vivo and, ultimately, in vivo natural-product glycorandomization.

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DO - 10.1016/j.chembiol.2005.04.009

M3 - Article

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AN - SCOPUS:24944496018

SN - 1074-5521

VL - 12

SP - 657

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JO - Chemistry and Biology

JF - Chemistry and Biology

IS - 6

ER -

Structure-based engineering of E. coli galactokinase as a first step toward in vivo glycorandomization

Abstract

Bibliographical note

Funding

ASJC Scopus subject areas

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