The EXT1/EXT2 tumor suppressors: catalytic activities and role in heparan sulfate biosynthesis.
The D-glucuronyltransferase and N-acetyl-D-glucosaminyltransferase reactions in heparan sulfate biosynthesis have been associated with two genes, EXT1 and EXT2, which are also implicated in the inherited bone disorder, multiple exostoses. Since the cell systems used to express recombinant EXT proteins synthesize endogenous heparan sulfate, and the EXT proteins tend to associate, it has not been possible to define the functional roles of the individual protein species. We therefore expressed EXT1 and EXT2 in yeast, which does not synthesize heparan sulfate. The recombinant EXT1 and EXT2 were both found to catalyze both glycosyltransferase reactions in vitro. Coexpression of the two proteins, but not mixing of separately expressed recombinant EXT1 and EXT2, yields hetero-oligomeric complexes in yeast and mammalian cells, with augmented glycosyltransferase activities. This stimulation does not depend on the membrane-bound state of the proteins.
Department of Medical Biochemistry and Microbiology, University of Uppsala, The Biomedical Center, Sweden.
C
C
Senay
T
T
Lind
K
K
Muguruma
Y
Y
Tone
H
H
Kitagawa
K
K
Sugahara
K
K
Lidholt
U
U
Lindahl
M
M
Kusche-Gullberg
D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002384:Catalysis; D005097:Exostoses, Multiple Hereditary / Q000201:enzymology / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D006497:Heparan Sulfate / Q000096:biosynthesis*; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics; D011233:Precipitin Tests; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D014170:Transformation, Genetic; D000099065:Exostosin 2; D000099058:Exostosin 1
D014453:Glucuronosyltransferase; D006497:Heparan Sulfate; D006596:Hexosaminidases; D008565:Membrane Proteins; D017351:N-Acetylglucosaminyltransferases; D011506:Proteins; D011994:Recombinant Proteins; D000099065:Exostosin 2; D000099058:Exostosin 1