Effects of primer-concentration on uronosyl-epimerization and sulfation patterns in p-hydroxyphenyl-O-beta-D-xylopyranoside-primed galactosaminoglycans produced by skin fibroblasts.
By supplying skin fibroblasts with different concentrations of the galactosaminoglycan chain-primer p-hydroxyphenyl-O-beta-D-xylopyranoside we have produced and recovered glycan-chains that were subsequently radio-iodinated in the hydroxyphenyl group and subjected to sequence analysis by using graded enzymic treatment followed by a combination of gel chromatography and electrophoresis. Fragments extending from the tagged reducing end to the cleavage-point were identified and quantified. Degradation by chondroitin B lyase of chains primed at 0.1 or 0.5 mM xyloside gave profiles indicating a periodic and wave-like distribution of iduronate-containing repeats, with high incidence around positions 2, 5 and onwards, whereas in chains produced at 1.0 mM xyloside the incidence of iduronate was similar in positions 1-4 and then declined. Degradation by chondroitin AC lyase indicated a high incidence of glucuronate in or near the linkage-region. There was a relatively uniform degree of sulfation in chains primed at low xyloside concentration, whereas chains primed at 1.0 mM xyloside gave very heterogeneous charge-patterns in all segments of the chain, including the linkage-region, giving the impression that adequate sulfation, probably at C-4 and at the first opportunity, is necessary to obtain an ordered and periodic epimerization pattern.
Cheng
F
F
Department of Cell and Molecular Biology, Lund University, Sweden.
Havsmark
B
B
Sakurai
K
K
Habuchi
H
H
Suzuki
S
S
Yoshida
K
K
Fransson
L A
LA
D002240:Carbohydrate Sequence; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D003871:Dermatan Sulfate / Q000737:chemistry / Q000378:metabolism; D004187:Disaccharides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D005347:Fibroblasts / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D005690:Galactose / Q000737:chemistry / Q000378:metabolism; D006027:Glycosides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008190:Lyases / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism*; D012867:Skin / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013431:Sulfates / Q000737:chemistry; D014316:Tritium; D014574:Uronic Acids / Q000737:chemistry*
D004187:Disaccharides; D006027:Glycosides; D009844:Oligosaccharides; D011134:Polysaccharides; D013431:Sulfates; D014574:Uronic Acids; C010807:xylosides; D014316:Tritium; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates; C007815:galactosaminoglycan; D008190:Lyases; D005690:Galactose