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. 1991 Aug 1;277(Pt 3):787–793. doi: 10.1042/bj2770787

Formation of two species of nascent proteochondroitin in separate loci of a microsomal preparation from chick-embryo epiphyseal cartilage.

G Sugumaran 1, J E Silbert 1
PMCID: PMC1151313  PMID: 1651703

Abstract

The potential relationship of an intact membrane organization to the synthesis of chondroitin was examined before and after modification of a chick-embryo cartilage microsomal system with the non-ionic detergent Triton X-100. Incubations with labelled UDP-GlcA and UDP-GalNAc indicated that Triton X-100 had little effect on the amount of chondroitin synthesized to form one species of large proteochondroitin (Type I). However, Triton X-100 had a marked stimulatory effect on the formation of another smaller species of proteochondroitin (Type II). Presence of this detergent during chondroitin polymerization also resulted in chains that were slightly smaller. Neither of the two proteochondroitin species were collagenase-sensitive, nor did they contain dermatan-like regions. Thus in these respects they were unlike the small proteochondroitins (PG-Lb or PG-Lt) that have been found in chick-embryo cartilage. They also differed greatly in size from these small proteoglycans as well as from the large aggregatable proteochondroitin (PG-H) from the same source. Synthesis of the larger (Type I) proteochondroitin species was not affected by prior treatment of the microsomes with chondroitin ABC lyase at concentrations sufficient for elimination of synthesis of most of the smaller (Type II) proteochondroitin species. Use of chondroitin ABC lyase subsequent to synthesis of the chondroitin also resulted in preferential degradation of the smaller species. Thus there were differences in formation and limitation in access of the chondroitin ABC lyase to the two species, consistent with other differences described previously. These results indicate that there are separate loci within the microsomal membranes for synthesis of the two species.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Faltynek C. R., Silbert J. E. Biosynthesis of chondroitin sulfate. Proteoglycans at the microsomal site of glycosaminoglycan formation. J Biol Chem. 1981 Jul 25;256(14):7203–7206. [PubMed] [Google Scholar]
  2. Habuchi H., Kimata K., Suzuki S. Changes in proteoglycan composition during development of rat skin. The occurrence in fetal skin of a chondroitin sulfate proteoglycan with high turnover rate. J Biol Chem. 1986 Jan 25;261(3):1031–1040. [PubMed] [Google Scholar]
  3. Kimura J. H., Thonar E. J., Hascall V. C., Reiner A., Poole A. R. Identification of core protein, an intermediate in proteoglycan biosynthesis in cultured chondrocytes from the Swarm rat chondrosarcoma. J Biol Chem. 1981 Aug 10;256(15):7890–7897. [PubMed] [Google Scholar]
  4. Lewis R. G., Spencer A. F., Silbert J. E. Biosynthesis of glycosoaminoglycans by microsomal preparations from cultured mastocytoma cells. Biochem J. 1973 Jun;134(2):465–471. doi: 10.1042/bj1340465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Noro A., Kimata K., Oike Y., Shinomura T., Maeda N., Yano S., Takahashi N., Suzuki S. Isolation and characterization of a third proteoglycan (PG-Lt) from chick embryo cartilage which contains disulfide-bonded collagenous polypeptide. J Biol Chem. 1983 Aug 10;258(15):9323–9331. [PubMed] [Google Scholar]
  6. Richmond M. E., DeLuca S., Silbert J. E. Biosynthesis of chondroitin sulfate. Assembly of chondroitin on microsomal primers. Biochemistry. 1973 Sep 25;12(20):3904–3910. doi: 10.1021/bi00744a018. [DOI] [PubMed] [Google Scholar]
  7. Richmond M. E., DeLuca S., Silbert J. E. Biosynthesis of chondroitin sulfate. Microsomal acceptors of sulfate, glucuronic acid, and N-acetylgalactosamine. Biochemistry. 1973 Sep 25;12(20):3898–3903. doi: 10.1021/bi00744a017. [DOI] [PubMed] [Google Scholar]
  8. SILBERT J. E. INCORPORATION OF 14C AND 3H FROM LABELED NUCLEOTIDE SUGARS INTO A POLYSACCHARIDE IN THE PRESENCE OF A CELL-FREE PREPARATION FROM CARTILAGE. J Biol Chem. 1964 May;239:1310–1315. [PubMed] [Google Scholar]
  9. Saito H., Yamagata T., Suzuki S. Enzymatic methods for the determination of small quantities of isomeric chondroitin sulfates. J Biol Chem. 1968 Apr 10;243(7):1536–1542. [PubMed] [Google Scholar]
  10. Shinomura T., Kimata K., Oike Y., Noro A., Hirose N., Tanabe K., Suzuki S. The occurrence of three different proteoglycan species in chick embryo cartilage. Isolation and characterization of a second proteoglycan (PG-Lb) and its precursor form. J Biol Chem. 1983 Aug 10;258(15):9314–9322. [PubMed] [Google Scholar]
  11. Silbert J. E. Biosynthesis of chondroitin sulfate. Chain termination. J Biol Chem. 1978 Oct 10;253(19):6888–6892. [PubMed] [Google Scholar]
  12. Silbert J. E., Palmer M. E., Humphries D. E., Silbert C. K. Formation of dermatan sulfate by cultured human skin fibroblasts. Effects of sulfate concentration on proportions of dermatan/chondroitin. J Biol Chem. 1986 Oct 15;261(29):13397–13400. [PubMed] [Google Scholar]
  13. Silbert J. E., Reppucci A. C., Jr Biosynthesis of chondroitin sulfate. Independent addition of glucuronic acid and N-acetylgalactosamine to oligosaccharides. J Biol Chem. 1976 Jul 10;251(13):3942–3947. [PubMed] [Google Scholar]
  14. Sugumaran G., Pisoni R. L., Silbert J. E. Biosynthesis of proteochondroitin sulfate: substrate requirements for formation of two independent species. Carbohydr Res. 1986 Aug 15;151:185–195. doi: 10.1016/s0008-6215(00)90339-2. [DOI] [PubMed] [Google Scholar]
  15. Sugumaran G., Silbert J. E. Biosynthesis of chondroitin sulfate. Organization of sulfation. J Biol Chem. 1989 Mar 5;264(7):3864–3868. [PubMed] [Google Scholar]
  16. Sugumaran G., Silbert J. E. Sulfation of chondroitin. Specificity, degree of sulfation, and detergent effects with 4-sulfating and 6-sulfating microsomal systems. J Biol Chem. 1988 Apr 5;263(10):4673–4678. [PubMed] [Google Scholar]

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