Molecular characterization of chicken syndecan-2 proteoglycan

LG Chen, J R Couchman, J Smith, A Woods*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

A partial syndecan-2 sequence (147 bp) was obtained from chicken embryonic fibroblast poly(A)(+) RNA by reverse transcription-PCR. This partial sequence was used to produce a 5'-end-labelled probe. A chicken liver cDNA library was screened with this probe, and overlapping clones were obtained encompassing the entire cDNA of 3 kb. The open reading frame encodes a protein of 201 amino acids. The cytoplasmic domain is identical with that of mammalian syndecan-2, and highly similar to those of Xenopus laevis and zebrafish syndecan-2. The transmembrane domain is identical with that of mammalian and zebrafish syndecan-2, and highly similar to that of Xenopus laevis syndecan-2. The ectodomain is 45-62% identical with that of zebrafish, Xenopus laevis and mammalian syndecan-2. Two coding single nucleotide polymorphisms were observed. In vitro transcription and translation yielded a product of 30 kDa. Western blotting of chicken embryonic fibroblast cell lysates with species-specific monoclonal antibody mAb 8.1 showed that chicken syndecan-2 is substituted with heparan sulphate, and that the major form of chicken syndecan-2 isolated from chicken fibroblasts is consistent with the formation of SIDS-resistant dimers, which is common for syndecans. A 5'-end-labelled probe hybridized to two mRNA species in chicken embryonic fibroblasts, while Northern analysis with poly(A)(+) RNAs from different tissues of chicken embryos showed wide and distinct distributions of chicken syndecan-2 during embryonic development. This pattern was different from that reported for syndecan-4, but consistent with the roles of syndecan-2 in neural maturation and in mesenchymal matrix interactions.

Original languageEnglish
Pages (from-to)481-490
Number of pages10
JournalBiochemical Journal
Volume366
DOIs
Publication statusPublished - 1 Sept 2002

Keywords / Materials (for Non-textual outputs)

  • BINDING DOMAIN
  • CELL-ADHESION
  • HEPARAN-SULFATE PROTEOGLYCANS
  • heparan sulphate
  • glycosaminoglycan
  • CYTOPLASMIC DOMAIN
  • embryonic development
  • PROTEIN-KINASE-C
  • DENDRITIC SPINES
  • FOCAL ADHESION FORMATION
  • PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE
  • EXPRESSION
  • CLONING

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