Combining carbochips and mass spectrometry to study the donor specificity for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase LgtA

Wanyi Guan; Lan Ban; Li Cai; Lei Li; Wenlan Chen; Xianwei Liu; Milan Mrksich; Peng George Wang

doi:10.1016/j.bmcl.2011.04.100

Combining carbochips and mass spectrometry to study the donor specificity for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase LgtA

Wanyi Guan, Lan Ban, Li Cai, Lei Li, Wenlan Chen, Xianwei Liu, Milan Mrksich, Peng George Wang^*

^*Corresponding author for this work

Biomedical Engineering

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

18 Scopus citations

Abstract

A library of 11 UDP-N-acetylglucosamine analogs were rapidly screened for their activities as donors for the Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA) by direct on-chip reaction and detection with SAMDI-TOF mass spectrometry. Six of the analogs were active in this assay and were analyzed by SAMDI to characterize the kinetics toward LgtA. The analysis revealed that substitutions on C-2, C-4, and C-6 affect the activity of the donors, with bulky groups at these positions decreasing affinity of the donors for the enzyme, and also revealed that activity is strongly affected by the stereochemistry at C-3, but not C-4, of the donor. The study is also significant because it demonstrates that SAMDI can be used to both profile glycosyltransferase activities and to provide a quantitative assessment of enzyme activity.

Original language	English (US)
Pages (from-to)	5025-5028
Number of pages	4
Journal	Bioorganic and Medicinal Chemistry Letters
Volume	21
Issue number	17
DOIs	https://doi.org/10.1016/j.bmcl.2011.04.100
State	Published - Sep 1 2011

Funding

This research was supported by the National Science Foundation ( CHE-0616892 to P.G.W., NSEC to M.M.), National Institute of Health ( R01 AI083754 to P.G.W. P50 GM086145 to M.M.), and National Cancer Institute ( R01 CA118208 to P.G.W.). W.G. acknowledges China Scholarship Council for financial support.

Keywords

Carbohydrate array
Glycosyltransferase
MALDI TOF MS
Unnatural sugar nucleotides

ASJC Scopus subject areas

Drug Discovery
Molecular Medicine
Molecular Biology
Biochemistry
Clinical Biochemistry
Pharmaceutical Science
Organic Chemistry

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10.1016/j.bmcl.2011.04.100

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title = "Combining carbochips and mass spectrometry to study the donor specificity for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase LgtA",

abstract = "A library of 11 UDP-N-acetylglucosamine analogs were rapidly screened for their activities as donors for the Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA) by direct on-chip reaction and detection with SAMDI-TOF mass spectrometry. Six of the analogs were active in this assay and were analyzed by SAMDI to characterize the kinetics toward LgtA. The analysis revealed that substitutions on C-2, C-4, and C-6 affect the activity of the donors, with bulky groups at these positions decreasing affinity of the donors for the enzyme, and also revealed that activity is strongly affected by the stereochemistry at C-3, but not C-4, of the donor. The study is also significant because it demonstrates that SAMDI can be used to both profile glycosyltransferase activities and to provide a quantitative assessment of enzyme activity.",

keywords = "Carbohydrate array, Glycosyltransferase, MALDI TOF MS, Unnatural sugar nucleotides",

author = "Wanyi Guan and Lan Ban and Li Cai and Lei Li and Wenlan Chen and Xianwei Liu and Milan Mrksich and Wang, {Peng George}",

note = "Funding Information: This research was supported by the National Science Foundation ( CHE-0616892 to P.G.W., NSEC to M.M.), National Institute of Health ( R01 AI083754 to P.G.W. P50 GM086145 to M.M.), and National Cancer Institute ( R01 CA118208 to P.G.W.). W.G. acknowledges China Scholarship Council for financial support. ",

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doi = "10.1016/j.bmcl.2011.04.100",

language = "English (US)",

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AU - Guan, Wanyi

AU - Ban, Lan

AU - Cai, Li

AU - Li, Lei

AU - Chen, Wenlan

AU - Liu, Xianwei

AU - Mrksich, Milan

AU - Wang, Peng George

N1 - Funding Information: This research was supported by the National Science Foundation ( CHE-0616892 to P.G.W., NSEC to M.M.), National Institute of Health ( R01 AI083754 to P.G.W. P50 GM086145 to M.M.), and National Cancer Institute ( R01 CA118208 to P.G.W.). W.G. acknowledges China Scholarship Council for financial support.

PY - 2011/9/1

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N2 - A library of 11 UDP-N-acetylglucosamine analogs were rapidly screened for their activities as donors for the Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA) by direct on-chip reaction and detection with SAMDI-TOF mass spectrometry. Six of the analogs were active in this assay and were analyzed by SAMDI to characterize the kinetics toward LgtA. The analysis revealed that substitutions on C-2, C-4, and C-6 affect the activity of the donors, with bulky groups at these positions decreasing affinity of the donors for the enzyme, and also revealed that activity is strongly affected by the stereochemistry at C-3, but not C-4, of the donor. The study is also significant because it demonstrates that SAMDI can be used to both profile glycosyltransferase activities and to provide a quantitative assessment of enzyme activity.

AB - A library of 11 UDP-N-acetylglucosamine analogs were rapidly screened for their activities as donors for the Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA) by direct on-chip reaction and detection with SAMDI-TOF mass spectrometry. Six of the analogs were active in this assay and were analyzed by SAMDI to characterize the kinetics toward LgtA. The analysis revealed that substitutions on C-2, C-4, and C-6 affect the activity of the donors, with bulky groups at these positions decreasing affinity of the donors for the enzyme, and also revealed that activity is strongly affected by the stereochemistry at C-3, but not C-4, of the donor. The study is also significant because it demonstrates that SAMDI can be used to both profile glycosyltransferase activities and to provide a quantitative assessment of enzyme activity.

KW - Carbohydrate array

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KW - MALDI TOF MS

KW - Unnatural sugar nucleotides

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Combining carbochips and mass spectrometry to study the donor specificity for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase LgtA

Abstract

Funding

Keywords

ASJC Scopus subject areas

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