Isolation of matrix vesicles by isoelectric focusing in Pevikon-Sephadex

S. Kakuta, D. Malamud, E. E. Golub, I. M. Shapiro

Research output: Contribution to journalArticlepeer-review

Abstract

We have investigated the use of an isoelectric focusing (IEF) technique for isolating and characterizing matrix vesicles. Focusing was performed on crude preparations of matrix vesicles isolated from collagenase digests of chick epiphyseal cartilage and purified by discontinuous sucrose gradient centrifugation. Crude and partially purified vesicle preparations were subjected to flat bed IEF in a slurry of Pevikon-Sephadex. Partially purified matrix vesicles focused as a narrow band (pl ∼- 6.5). Alkaline phosphatase, solubilized from matrix vesicles, focused with a pl of 4.0-4.5. The IEF profile of matrix vesicles also differed from that of chondrocyte membranes. Thus, the membrane pls were ∼- 5.4 and 6.6-7.8, respectively. The latter peak probably corresponded to the pl of the matrix vesicle preparation. This observation lends support to the view that vesicles originate from distinct regions of the chondrocyte membrane.

Original languageEnglish (US)
Pages (from-to)187-191
Number of pages5
JournalBone
Volume6
Issue number3
DOIs
StatePublished - 1985

Keywords

  • Calcification
  • Growth Cartilage
  • Isoelectric Focusing
  • Matrix Vesicles
  • Pevikon-Sephadex

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

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