Cloning and characterisation of the UDP-glucose 4 '-epimerase of Trypanosoma cruzi

Janine R.  Roper; Michael A. J.  Ferguson

doi:10.1016/j.molbiopara.2003.07.002

Cloning and characterisation of the UDP-glucose 4 '-epimerase of Trypanosoma cruzi

Janine R. Roper, Michael A. J. Ferguson

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27 Citations (Scopus)

Abstract

Trypanosoma cruzi incorporates galactose into many of its cell-surface glycoconjugates but it is unable to transport this sugar through its hexose transporter. Epimerisation of UDP-glucose to UDP-galactose by UDP-glucose 4'-epimerase may be the only way that the parasites can obtain galactose. Here, we describe cloning the T. cruzi UDP-Glc 4'-epimerase (TcGALE) gene and show that it is functional by complementing an Escherichia coli epimerase-deficient strain. The T. cruzi GALE gene encodes a 42.4 kDa protein and the recombinant protein expressed in E. coli is a homodimer in solution with a specific activity of 3.8 U mg(-1) and K-m for UDP-Gal of 114 µM. Unlike the human epimerase. T. cruzi UDP-Glc 4'-epimerase is unable to inter-convert UDP-N-acetylglucosamine and UDP-N-acetylgalactosamine. This may explain why T. cruzi initiates O-glycosylation of its abundant GPI-anchored surface mucins via GlcNAcalpha1-O-Thr/Ser rather than the GalNAcalpha1-O-Thr/Ser linkage that is common for mucins from many other eukaryotes. (C) 2003 Elsevier B.V. All rights reserved.

Original language	English
Pages (from-to)	47-53
Number of pages	7
Journal	Molecular and Biochemical Parasitology
Volume	132
Issue number	1
DOIs	https://doi.org/10.1016/j.molbiopara.2003.07.002
Publication status	Published - Nov 2003

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title = "Cloning and characterisation of the UDP-glucose 4 '-epimerase of Trypanosoma cruzi",

abstract = "Trypanosoma cruzi incorporates galactose into many of its cell-surface glycoconjugates but it is unable to transport this sugar through its hexose transporter. Epimerisation of UDP-glucose to UDP-galactose by UDP-glucose 4'-epimerase may be the only way that the parasites can obtain galactose. Here, we describe cloning the T. cruzi UDP-Glc 4'-epimerase (TcGALE) gene and show that it is functional by complementing an Escherichia coli epimerase-deficient strain. The T. cruzi GALE gene encodes a 42.4 kDa protein and the recombinant protein expressed in E. coli is a homodimer in solution with a specific activity of 3.8 U mg(-1) and K-m for UDP-Gal of 114 µM. Unlike the human epimerase. T. cruzi UDP-Glc 4'-epimerase is unable to inter-convert UDP-N-acetylglucosamine and UDP-N-acetylgalactosamine. This may explain why T. cruzi initiates O-glycosylation of its abundant GPI-anchored surface mucins via GlcNAcalpha1-O-Thr/Ser rather than the GalNAcalpha1-O-Thr/Ser linkage that is common for mucins from many other eukaryotes. (C) 2003 Elsevier B.V. All rights reserved.",

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AU - Ferguson, Michael A. J.

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N2 - Trypanosoma cruzi incorporates galactose into many of its cell-surface glycoconjugates but it is unable to transport this sugar through its hexose transporter. Epimerisation of UDP-glucose to UDP-galactose by UDP-glucose 4'-epimerase may be the only way that the parasites can obtain galactose. Here, we describe cloning the T. cruzi UDP-Glc 4'-epimerase (TcGALE) gene and show that it is functional by complementing an Escherichia coli epimerase-deficient strain. The T. cruzi GALE gene encodes a 42.4 kDa protein and the recombinant protein expressed in E. coli is a homodimer in solution with a specific activity of 3.8 U mg(-1) and K-m for UDP-Gal of 114 µM. Unlike the human epimerase. T. cruzi UDP-Glc 4'-epimerase is unable to inter-convert UDP-N-acetylglucosamine and UDP-N-acetylgalactosamine. This may explain why T. cruzi initiates O-glycosylation of its abundant GPI-anchored surface mucins via GlcNAcalpha1-O-Thr/Ser rather than the GalNAcalpha1-O-Thr/Ser linkage that is common for mucins from many other eukaryotes. (C) 2003 Elsevier B.V. All rights reserved.

AB - Trypanosoma cruzi incorporates galactose into many of its cell-surface glycoconjugates but it is unable to transport this sugar through its hexose transporter. Epimerisation of UDP-glucose to UDP-galactose by UDP-glucose 4'-epimerase may be the only way that the parasites can obtain galactose. Here, we describe cloning the T. cruzi UDP-Glc 4'-epimerase (TcGALE) gene and show that it is functional by complementing an Escherichia coli epimerase-deficient strain. The T. cruzi GALE gene encodes a 42.4 kDa protein and the recombinant protein expressed in E. coli is a homodimer in solution with a specific activity of 3.8 U mg(-1) and K-m for UDP-Gal of 114 µM. Unlike the human epimerase. T. cruzi UDP-Glc 4'-epimerase is unable to inter-convert UDP-N-acetylglucosamine and UDP-N-acetylgalactosamine. This may explain why T. cruzi initiates O-glycosylation of its abundant GPI-anchored surface mucins via GlcNAcalpha1-O-Thr/Ser rather than the GalNAcalpha1-O-Thr/Ser linkage that is common for mucins from many other eukaryotes. (C) 2003 Elsevier B.V. All rights reserved.

U2 - 10.1016/j.molbiopara.2003.07.002

DO - 10.1016/j.molbiopara.2003.07.002

M3 - Article

SN - 0166-6851

VL - 132

SP - 47

EP - 53

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

IS - 1

ER -

Cloning and characterisation of the UDP-glucose 4 '-epimerase of Trypanosoma cruzi

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