UDP-glucuronate decarboxylase, a key enzyme in proteoglycan synthesis: cloning, characterization, and localization.
UDP-glucuronate decarboxylase (UGD) catalyzes the formation of UDP-xylose from UDP-glucuronate. UDP-xylose is then used to initiate glycosaminoglycan biosynthesis on the core protein of proteoglycans. In a yeast two-hybrid screen with the protein kinase Akt (protein kinase B), we detected interactions with a novel sequence, which we cloned and expressed. The expressed protein displayed UGD activity but did not display the activities of homologous nucleotide sugar epimerases or dehydratases. We did not detect phosphorylation of UGD by Akt nor did we detect any influence of Akt on UGD activity. Effects of UGD on Akt kinase activity were also absent. Northern blot and Western blot analyses revealed the presence of UGD in multiple tissues and brain regions. Subcellular studies and histochemistry localized UGD protein to the perinuclear Golgi where xylosylation of proteoglycan core proteins is known to occur.
Moriarity
John L
JL
Department of Neurological Surgery, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.
Hurt
K Joseph
KJ
Resnick
Adam C
AC
Storm
Phillip B
PB
Laroy
Wouter
W
Schnaar
Ronald L
RL
Snyder
Solomon H
SH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002262:Carboxy-Lyases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D005982:Glutathione Transferase / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases; D011509:Proteoglycans / Q000737:chemistry; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D051381:Rats; D017386:Sequence Homology, Amino Acid; D021241:Spectrometry, Mass, Electrospray Ionization; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014534:UDPglucose 4-Epimerase / Q000378:metabolism
D018076:DNA, Complementary; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins; D005982:Glutathione Transferase; C494918:AKT1 protein, human; C494920:Akt1 protein, rat; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D002262:Carboxy-Lyases; C023257:UDPglucuronate decarboxylase; D014534:UDPglucose 4-Epimerase