JBC, Vol. 250, Issue 17, 6904-6915, Sep, 1975
Biosynthesis and characterization of lipid-linked sugars and glycoproteins in aorta
J. Chambers and A. D. Elbein
A particulate enzyme from bovine aorta catalyzes the incorporation of
mannose from GDP-D-[14C]mannose into three products as follows: 1. Most of
the radioactivity which is incorporated in short term incubations is into a
product that is soluble in CHCl3/CH3OH (2/1, v/v). This product was
purified by chromatography on DEAE-cellulose and Sephadex LH-20. The
purified glycolipid was stable to alkaline saponification but released
[14C]mannose when subjected to mild acid hydrolysis (1/2 = 7 min at 100
degrees in 0.01 N HCl). The purified glycolipid had the same mobility on
silica gel plates in an acidic, basic, or neutral solvent system as did
glycolipid had the same mobility on silica gel plates in an acidic, basic,
or neutral solvent system as did authentic dolichyl mannopyranosyl
phosphate. The synthesis of the 14C-mannolipid was reversed by the addition
of GDP and Mg2+. 2. [14C]mannose is also incorporated, although at a slower
rate into products which are soluble in CHCl3/CH3OH/H2O (1/1/0.3, v/v).
When the 1/10.3 soluble material was chromatographed on Avicel plates, it
gave rise to three distinct radioactive bands which appear to be
lipid-linked oligosaccharides. Mild acid hydrolysis of the 1/10.3 soluble
material released water-soluble, neutral 14C-oligosaccharides which eluted
from Sephadex G-50 in two or three peaks between the standards cytochrome c
and GDP-mannose...
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?