TY - JOUR
T1 - Reconstitution in vitro of the GDP-fucose biosynthetic pathways of Caenorhabditis elegans and Drosophila melanogaster
AU - Rhomberg, Simone
AU - Fuchsluger, Christina
AU - Rendić, Dubravko
AU - Paschinger, Katharina
AU - Jantsch, Verena
AU - Kosma, Paul
AU - Wilson, Iain B H
PY - 2006/5
Y1 - 2006/5
N2 - The deoxyhexose sugar fucose has an important fine-tuning role in regulating the functions of glycoconjugates in disease and development in mammals. The two genetic model organisms Caenorhabditis elegans and Drosophila melanogaster also express a range of fucosylated glycans, and the nematode particularly has a number of novel forms. For the synthesis of such glycans, the formation of GDP-fucose, which is generated from GDP-mannose in three steps catalysed by two enzymes, is required. By homology we have identified and cloned cDNAs encoding these two proteins, GDP-mannose dehydratase (GMD; EC 4.2.1.47) and GDP-keto-6-deoxymannose 3,5-epimerase/4-reductase (GER or FX protein; EC 1.1.1.271), from both Caenorhabditis and Drosophila. Whereas the nematode has two genes encoding forms of GMD (gmd-1 and gmd-2) and one GER-encoding gene (ger-1), the insect has, like mammalian species, only one homologue of each (gmd and gmer). This compares to the presence of two forms of both enzymes in Arabidopsis thaliana. All corresponding cDNAs from Caenorhabditis and Drosophila, as well as the previously uncharacterized Arabidopsis GER2, were separately expressed, and the encoded proteins found to have the predicted activity. The biochemical characterization of these enzymes is complementary to strategies aimed at manipulating the expression of fucosylated glycans in these organisms.
AB - The deoxyhexose sugar fucose has an important fine-tuning role in regulating the functions of glycoconjugates in disease and development in mammals. The two genetic model organisms Caenorhabditis elegans and Drosophila melanogaster also express a range of fucosylated glycans, and the nematode particularly has a number of novel forms. For the synthesis of such glycans, the formation of GDP-fucose, which is generated from GDP-mannose in three steps catalysed by two enzymes, is required. By homology we have identified and cloned cDNAs encoding these two proteins, GDP-mannose dehydratase (GMD; EC 4.2.1.47) and GDP-keto-6-deoxymannose 3,5-epimerase/4-reductase (GER or FX protein; EC 1.1.1.271), from both Caenorhabditis and Drosophila. Whereas the nematode has two genes encoding forms of GMD (gmd-1 and gmd-2) and one GER-encoding gene (ger-1), the insect has, like mammalian species, only one homologue of each (gmd and gmer). This compares to the presence of two forms of both enzymes in Arabidopsis thaliana. All corresponding cDNAs from Caenorhabditis and Drosophila, as well as the previously uncharacterized Arabidopsis GER2, were separately expressed, and the encoded proteins found to have the predicted activity. The biochemical characterization of these enzymes is complementary to strategies aimed at manipulating the expression of fucosylated glycans in these organisms.
KW - Amino Acid Sequence
KW - Animals
KW - Arabidopsis
KW - Arabidopsis Proteins
KW - Caenorhabditis elegans
KW - Caenorhabditis elegans Proteins
KW - Carbohydrate Epimerases
KW - Cloning, Molecular
KW - DNA, Complementary
KW - Drosophila Proteins
KW - Drosophila melanogaster
KW - Gene Expression Regulation, Developmental
KW - Guanosine Diphosphate Fucose
KW - Guanosine Diphosphate Mannose
KW - Humans
KW - Hydro-Lyases
KW - Molecular Sequence Data
KW - Protein Isoforms
KW - Sequence Alignment
KW - Sequence Homology, Amino Acid
KW - Species Specificity
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1111/j.1742-4658.2006.05239.x
DO - 10.1111/j.1742-4658.2006.05239.x
M3 - Article
C2 - 16650000
SN - 1742-464X
VL - 273
SP - 2244
EP - 2256
JO - The FEBS Journal
JF - The FEBS Journal
IS - 10
ER -