Identification of a UDP-Gal: GlcNAc-R galactosyltransferase activity in Escherichia coli VW187

Pedro J Montoya-Peleaz; John G Riley; Walter A Szarek; Miguel A Valvano; John S Schutzbach; Inka Brockhausen

doi:10.1016/j.bmcl.2004.11.077

Identification of a UDP-Gal: GlcNAc-R galactosyltransferase activity in Escherichia coli VW187

Pedro J Montoya-Peleaz, John G Riley, Walter A Szarek, Miguel A Valvano, John S Schutzbach, Inka Brockhausen

School of Medicine, Dentistry and Biomedical Sciences

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

39 Citations (Scopus)

Abstract

A novel acceptor substrate for galactosyltransferase was synthesized containing GlcNAcalpha-pyrophosphate, covalently bound to a hydrophobic phenoxyundecyl moiety (GlcNAc alpha-O-PO(3)-PO(3)-(CH(2))(11)-O-Phenyl). The new substrate was used to develop an assay for a galactosyltransferase activity from Escherichia coli strain VW187 that is involved in lipopolysaccharide synthesis and has not been studied by others. We showed that Gal was transferred from UDP-Gal to the novel acceptor substrate. This was a significant improvement over our previous preliminary assays of the enzyme using endogenous substrate, and showed that these synthetic substrates are useful for assaying enzymes that utilize lipid-bound substrates in O-chain synthesis in Gram-negative bacteria.

Original language	English
Pages (from-to)	1205-11
Number of pages	7
Journal	Bioorganic & Medicinal Chemistry Letters
Volume	15
Issue number	4
DOIs	https://doi.org/10.1016/j.bmcl.2004.11.077
Publication status	Published - 15 Feb 2005

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Organic Chemistry
Drug Discovery
Pharmaceutical Science

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title = "Identification of a UDP-Gal: GlcNAc-R galactosyltransferase activity in Escherichia coli VW187",

abstract = "A novel acceptor substrate for galactosyltransferase was synthesized containing GlcNAcalpha-pyrophosphate, covalently bound to a hydrophobic phenoxyundecyl moiety (GlcNAc alpha-O-PO(3)-PO(3)-(CH(2))(11)-O-Phenyl). The new substrate was used to develop an assay for a galactosyltransferase activity from Escherichia coli strain VW187 that is involved in lipopolysaccharide synthesis and has not been studied by others. We showed that Gal was transferred from UDP-Gal to the novel acceptor substrate. This was a significant improvement over our previous preliminary assays of the enzyme using endogenous substrate, and showed that these synthetic substrates are useful for assaying enzymes that utilize lipid-bound substrates in O-chain synthesis in Gram-negative bacteria.",

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T2 - GlcNAc-R galactosyltransferase activity in Escherichia coli VW187

AU - Montoya-Peleaz, Pedro J

AU - Riley, John G

AU - Szarek, Walter A

AU - Valvano, Miguel A

AU - Schutzbach, John S

AU - Brockhausen, Inka

PY - 2005/2/15

Y1 - 2005/2/15

N2 - A novel acceptor substrate for galactosyltransferase was synthesized containing GlcNAcalpha-pyrophosphate, covalently bound to a hydrophobic phenoxyundecyl moiety (GlcNAc alpha-O-PO(3)-PO(3)-(CH(2))(11)-O-Phenyl). The new substrate was used to develop an assay for a galactosyltransferase activity from Escherichia coli strain VW187 that is involved in lipopolysaccharide synthesis and has not been studied by others. We showed that Gal was transferred from UDP-Gal to the novel acceptor substrate. This was a significant improvement over our previous preliminary assays of the enzyme using endogenous substrate, and showed that these synthetic substrates are useful for assaying enzymes that utilize lipid-bound substrates in O-chain synthesis in Gram-negative bacteria.

AB - A novel acceptor substrate for galactosyltransferase was synthesized containing GlcNAcalpha-pyrophosphate, covalently bound to a hydrophobic phenoxyundecyl moiety (GlcNAc alpha-O-PO(3)-PO(3)-(CH(2))(11)-O-Phenyl). The new substrate was used to develop an assay for a galactosyltransferase activity from Escherichia coli strain VW187 that is involved in lipopolysaccharide synthesis and has not been studied by others. We showed that Gal was transferred from UDP-Gal to the novel acceptor substrate. This was a significant improvement over our previous preliminary assays of the enzyme using endogenous substrate, and showed that these synthetic substrates are useful for assaying enzymes that utilize lipid-bound substrates in O-chain synthesis in Gram-negative bacteria.

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DO - 10.1016/j.bmcl.2004.11.077

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SN - 0960-894X

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JO - Bioorganic & Medicinal Chemistry Letters

JF - Bioorganic & Medicinal Chemistry Letters

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ER -

Identification of a UDP-Gal: GlcNAc-R galactosyltransferase activity in Escherichia coli VW187

Abstract

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