Electron-microscopic and electrophoretic studies of bovine femoral-head cartilage proteoglycan fractions

D. J. Thornton, I. A. Nieduszynski, K. Oates, J .K. Sheehan

Research output: Contribution to journalArticlepeer-review

Abstract

Proteoglycans (A1D1) extracted from bovine femoral-head cartilage were examined by electron microscopy using benzyldimethylammonium chloride as a spreading agent. The preparation contained a mixture of particles, some with a 'beaded' structure and a contiguous filamentous 'tail' at one end and others which appeared as round 'blobs', some of which also had filamentous tails. Previous electron-microscopic studies of proteoglycan monomers have indicated that their length distributions were apparently unimodal, a finding that contrasted with agarose/polyacrylamide-gel-electrophoresis results, which generally indicated two bands. In the present study proteoglycans isolated from the slowly migrating electrophoretic band were shown to be predominantly the larger molecules of beaded appearance, whereas the rapidly migrating proteoglycans were predominantly molecules with the 'blob-like' appearance. Gel-filtration, isopycnic-density-gradient-centrifugation and rate-zonal-centrifugation techniques were evaluated as means of proteoglycan fractionation by electron microscopy and agarose-gel electrophoresis. Rate-zonal centrifugation in mixed-salt gradients of caesium chloride/4 M-guanidinium chloride yielded the most effective fractionation.

Original languageEnglish
Pages (from-to)41-48
Number of pages8
JournalThe Biochemical Journal
Volume240
Issue number1
Publication statusPublished - 15 Nov 1986

Keywords

  • Animals
  • Cartilage, Articular/analysis
  • Cattle
  • Centrifugation, Zonal
  • Chromatography, Gel
  • Electrophoresis, Agar Gel
  • Femur Head/analysis
  • Microscopy, Electron
  • Proteoglycans/analysis

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