caa a22 4500 469114894 CHVBK 20180323133122.0 cr unu---uuuuu 170328e19920201xx s 000 0 eng 10.1007/BF00731177 doi (NATIONALLICENCE)springer-10.1007/BF00731177 Sequence analysis of p -hydroxyphenyl- O -β- d -xyloside initiated and radio-iodinated dermatan sulfate from skin fibroblasts [Elektronische Daten] [Lars-Åke Fransson, Birgitta Havsmark, Katsukiyo Sakurai, Sakaru Suzuki] To generate xyloside-primed dermatan sulfate suitable for sequence analysis, skin fibroblasts were incubated withp-hydroxyphenyl-β-d-xylopyranoside and [3H]galactose, and free [3H]glycosaminoglycan chains were isolated from the culture medium by ion exchange and gel chromatography. After125I labelling of their reducing-terminal hydroxyphenyl groups, chains were subjected to various chemical and enzymatic degradations, both partial and complete, followed by gradient polyacrylamide gel electrophoresis and autoradiographic identification of fragments extending from the labelled reducing-end to the point of cleavage. Results of periodate oxidation-alkaline scission indicated that the xylose moiety remained unsubstituted at C-2/C-3; exhaustive treatment with chondroitin AC-I lyase afforded the fragment ΔHexA-Gal-Gal-Xyl-R (R = radio-iodinated hydroxyphenyl group), and complete degradations with chondroitin ABC lyase as well as testicular hyaluronidase yielded the fragments ΔHexA/HexA-GalNAc-GlcA-Gal-Gal-Xyl-R with or without sulfate on theN-acetylgalactosamine. Partial digestions with testicular hyaluronidase or chondroitin B lyase indicated that glucuronic acid was common in the first three repeats after the linkage region and that iduronic acid could occupy any position thereafter. Hence, there were no indications of a repeated, periodic appearance of the clustered GlcA-GalNAc repeats which was previously observed in proteoglycan derived dermatan sulfate [Fransson L-Å, Havsmark B, Silverberg I (1990)Biochem J 269:381-8], suggesting a role for the protein part in controlling the formation of particular copolymeric features during glycosaminoglycan assembly. Chapman and Hall, 1992 glycosaminoglycan nationallicence dermatan sulfate nationallicence xylosides nationallicence sequencing nationallicence GAG : glycosaminoglycans nationallicence CS : chondroitin sulfate nationallicence DS : dermatan sulfate nationallicence Ser : serine nationallicence Xyl : d-xylose nationallicence Gal : d-galactose nationallicence GlcA : d-glucuronic acid nationallicence IdoA : l-iduronic acid nationallicence GalNAc : N-acetyl-d-galactosamine nationallicence GlcNAc : N-acetyl-d-glucosamine nationallicence ΔHexA : 4-deoxy-l-threo-hex-4-enopyranosyluronic acid nationallicence HO-Phe : p-hydroxyphenyl group nationallicence HO-Phe-Xyl : p-hydroxyphenyl-O-β-d-xylopyranoside nationallicence O2N-Phe-Xyl : p-nitrophenyl-β-d-xylopyranoside nationallicence OSO3 : ester sulfate nationallicence PAGE : polyacrylamide gel electrophoresis nationallicence HPLC : high performance liquid chromatography nationallicence FPLC : fast performance liquid chromatography nationallicence LC : standard liquid chromatography nationallicence Fransson Lars-Åke Department of Medical and Physiological Chemistry, University of Lund, P.O. Box 94, S-221 00, Lund, Sweden aut Havsmark Birgitta Department of Medical and Physiological Chemistry, University of Lund, P.O. Box 94, S-221 00, Lund, Sweden aut Sakurai Katsukiyo Seikagaku Corporation, Tokyo Research Institute, 189, Tokyo, Japan aut Suzuki Sakaru Institute for Molecular Science of Medicine, Aichi Medical University, 480-11, Nagakute, Aichi, Japan aut Glycoconjugate Journal Kluwer Academic Publishers 9/1(1992-02-01), 45-55 0282-0080 9:1<45 1992 9 10719 https://doi.org/10.1007/BF00731177 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00731177 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fransson Lars-Åke Department of Medical and Physiological Chemistry, University of Lund, P.O. Box 94, S-221 00, Lund, Sweden aut NATIONALLICENCE 700 1- Havsmark Birgitta Department of Medical and Physiological Chemistry, University of Lund, P.O. Box 94, S-221 00, Lund, Sweden aut NATIONALLICENCE 700 1- Sakurai Katsukiyo Seikagaku Corporation, Tokyo Research Institute, 189, Tokyo, Japan aut NATIONALLICENCE 700 1- Suzuki Sakaru Institute for Molecular Science of Medicine, Aichi Medical University, 480-11, Nagakute, Aichi, Japan aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 9/1(1992-02-01), 45-55 0282-0080 9:1<45 1992 9 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer