NDLI: Purification and characterization of chick corneal beta-D-glucuronyltransferase involved in chondroitin sulfate biosynthesis.

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Purification and characterization of chick corneal beta-D-glucuronyltransferase involved in chondroitin sulfate biosynthesis.

Content Provider	Semantic Scholar
Author	Ogata, Nana Nakazawa, Kiyoshi
Copyright Year	2002
Abstract	Beta-D-Glucuronyltransferase, which transfers D-glucuronic acid (GlcA) from UDP-GlcA to N-acetyl-D-galactosamine (GalNAc) at the nonreducing end of chondro-pentasaccharide-PA (pyridylamino-), GalNAcbeta1-(4GlcAbeta1-3GalNAcbeta1)2-PA, was purified 339-fold with an 11.0% yield from 2-d-old chick corneas by chromatography on DEAE-Sepharose, WGA-agarose, heparin-Sepharose, and 1st and 2nd UDP-GlcA-agarose (in the presence of Gal) columns. The activity was detected by fluorescence of PA residues of the product. The purified enzyme has an optimum pH of 7.0 (Mes buffer), and much higher activity toward chondro-heptasaccharide-PA than toward the chondro-pentasaccharide-PA, but no activity toward p-nitrophenyl-beta-GalNAc. The enzyme activity was almost completely inhibited by GalNAc (20 mm). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme fraction showed one band of 38 kDa with many other bands. The amino acid sequence was determined for the tryptic digests of the 38 kDa band protein. The sequences determined showed no homology to those of several beta-glucuronyltransferases reported previously. It seems that the enzyme is involved in the elongation of chondroitin sulfate chains in vivo.
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://bpb.pharm.or.jp/bpb/200210/b10_1282.pdf
PubMed reference number	12392079v1
Volume Number	25
Issue Number	10
Journal	Biological & pharmaceutical bulletin
Language	English
Access Restriction	Open
Subject Keyword	Acetylcysteine Amino Acid Sequence Amino Acids Anabolism Bands Buffers Chondroitin Sulfates Chromatography, DEAE-Cellulose Cornea Fluorescence Galactosamine Gel Electrophoresis (lab technique) Glucuronic Acid Glucuronosyltransferase Homologous Gene Polyacrylamide Gel Electrophoresis SDS-PAGE Sodium Dodecyl Sulfate Thioctic Acid Uridine Diphosphate Whole Genome Amplification beta Thalassemia chondroitin sulfate biosynthetic process enzyme activity heparin polyacrylamide gels
Content Type	Text
Resource Type	Article

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