Porcine SPARC: Isolation from dentin, cDNA sequence, and computer model

Yong Hee P. Chun, Yasuo Yamakoshi, Jung Wook Kim, Takanori Iwata, Jan C.C. Hu, James P. Simmer

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Genes encoding the major enamel matrix proteins and non-collagenous proteins of bone and dentin are members of the secretory calcium-binding phosphoprotein (SCPP) family, which originated from ancestral SPARC (secreted protein, acidic and rich in cysteine; BM-40/osteonectin). To better understand the role of SPARC in mineralizing systems, we isolated SPARC from developing pig teeth, deduced its primary structure from the cDNA sequence, and determined its quaternary structure by homology modelling with reference to human SPARC crystal structures. The guanidine/EDTA extract from porcine dentin was fractionated by anion-exchange and size-exclusion chromatography. Stains-all positive bands at 38 and 35 kDa gave the N-terminal sequences APQQEALPDETEV and DFEKNYNMYIFPV, which corresponded to the SPARC N terminus and an internal region of the protein. Porcine SPARC contains 300 amino acids, including the 17-amino acid signal peptide, and shares 96.2% amino acid sequence identity with human SPARC. Without post-translational modifications, the 283-amino acid secreted protein has a molecular mass of 32.3 kDa. The three-dimensional model revealed that porcine SPARC contains a single N-linked glycosylation at N113, seven intramolecular disulfide bridges, and assembles into dimers. SPARC is composed of three structural/functional domains: an acidic Ca2+-binding, a follistatin-like, and an extracellular calcium-binding domain.

Original languageEnglish (US)
Pages (from-to)78-85
Number of pages8
JournalEuropean Journal of Oral Sciences
Volume114
Issue numberSUPPL. 1
DOIs
StatePublished - May 1 2006
Externally publishedYes

Keywords

  • BM-40
  • Dentin
  • Enamel
  • Follistatin
  • Osteonectin
  • SPARC

ASJC Scopus subject areas

  • Dentistry(all)

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