Isolation and some structure analyses of a copolymeric chondroitin sulfate-dermatan sulfate proteoglycan from post-burn, human hypertrophic scar.
A D-glucuronic acid rich, copolymeric chondroitin sulfate (CS)-dermatan sulfate (DS) proteoglycan (PG) from post-burn hypertrophic scar tissue (HSc) was obtained by DEAE-cellulose chromatography and differential ethanol fractionation, and further purified on a Sepharose CL-6B column. CS-DS-PG protein content was 14% (w/w). The amino-terminal amino acid sequence of the first ten residues was as follows: NH2-Asp-Glu-Ala-B-Gly-Ile-Gly-Pro-Glu-Val. This sequence is identical to that of human embryonic fibroblast cell (IMR-90) CS-DS-PG, as well as to human HSc-DS-PG. After chondroitinase ABC treatment, two peptides (Mr 22,000 and 16,000 daltons) were detected by sodium dodecyl sulfate-(polyacryl)amide gel electrophoresis (SDS-PAGE). ELISA analysis using rabbit antiserum raised against a synthetic peptide that contained 15 amino acids in the same sequence as the amino terminus of human fetal membrane PG showed significant reactivity with HSc CS-DS-PG. HSc CS-DS-PG had an apparent Mr of approximately 78,000 daltons, as determined by Sepharose CL-6B chromatography and SDS-PAGE. Alkaline borohydride treatment of CS-DS-PG liberated CS-DS glycosaminoglycan (GAG) chains having an Mr of 29,000 daltons. The conversion of xylose to xylitol indicated that the GAG chains are attached to the PG protein core at O-3 through a xylosyl-seryl linkage. CS-DS-PG also contained both N and O-linked oligosaccharides and did not aggregate with hyaluronic acid. These results, together with those reported previously, showed that HSc CS-DS-PG and DS-PG have the same A1-A15 amino acid sequence at the amino terminus but different protein cores. HSc CS-DS-PG was completely digested with chondroitinase AC and is, therefore, distinctly different from HSc DS-PG.
Garg
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Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, at Shriners Burns Institute, Boston, Massachusetts 02114.
Siebert
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Swann
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D053674:Aggrecans; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002056:Burns / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis; D002807:Chondroitin / Q000031:analogs & derivatives*; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D002921:Cicatrix / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000032:analysis*; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000032:analysis*; D012867:Skin / Q000032:analysis
D053674:Aggrecans; D000596:Amino Acids; D002241:Carbohydrates; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D003871:Dermatan Sulfate; D002807:Chondroitin