Partial purification of xylosyltransferase (XylT) from rat liver and characterization of endogenous acceptors

Conference · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6985992

Klinger, M; Roden, L

The biosynthesis of the carbohydrate-protein linkage region of most proteoglycan species is initiated by transfer of xylose from UDP-xylose to serine hydroxyl groups in the core protein. The XylT catalyzing this reaction has been previously purified from embryonic chick cartilage and from a rat chondrosarcoma but not from a normal mammalian tissue. In this study, XylT was extracted from rat liver by homogenization in buffer containing 1 M KCl and was partially purified by chromatography on heparin-Sepharose, AH-Sepharose, and on Sepharose-linked tryptic fragments of silk fibroin. The eluate from the latter contained more than 40% of the applied activity and less than 5% of the protein. Gel chromatography of XylT eluted from heparin-Sepharose indicated a mol. wt. of 95,000 to 100,000. Incorporation of (/sup 3/H)xylose into endogenous acceptors in the crude extract amounted to more than 50% of the total observed with added substrate (silk fibroin). Of the total endogenous acceptor activity in the crude extract, 98% was not adsorbed to heparin-Sepharose and yielded a labeled product which was stable to treatment with 0.5 M NaOH at 20/sup 0/C for 16 h; this material may have been glycogen. In contrast, most of the radioactivity incorporated into the endogenous acceptor in the heparin-Sepharose eluate was alkali-labile, as would be expected for the xylosylated core protein of a proteoglycan.

• Research Organization: Univ. of Alabama, Birmingham
• OSTI ID: 6985992
• Report Number(s): CONF-8606151-; TRN: 86-039075
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
TRANSFERASES
PURIFICATION
CHROMATOGRAPHY
GLUCOPROTEINS
LIVER
MOLECULAR WEIGHT
RATS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
XYLOSE
ALDEHYDES
ANIMALS
BODY
CARBOHYDRATES
DIGESTIVE SYSTEM
ENZYMES
GLANDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MONOSACCHARIDES
ORGANIC COMPOUNDS
ORGANS
PENTOSES
PROTEINS
RODENTS
SACCHARIDES
SEPARATION PROCESSES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques