Further application of a two-step heparin affinity chromatography method using divalent cations as eluents: Purification and identification of membrane-bound heparin binding proteins from the mitochondrial fraction of HL-60 cells

Tsukimi Iida, Masaharu Kamo, Nobuyuki Uozumi, Takashi Inui, Katsuyuki Imai

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Membrane proteins were obtained from the mitochondrial fraction of HL-60 cells by solubilization with octyl glucoside and bound to heparin-gels. Bound proteins were successively eluted with solutions containing increasing concentrations of Mg2+ in the first and increasing concentrations of Ca2+ in the second chromatography. After SDS-PAGE and subsequent N-terminal amino acid analysis of proteins on each band, 13 proteins were identified. Fifteen out of the 37 proteins analysed were modified at their N-termini. These results show that this two-step affinity chromatography method using divalent cations as eluents can be applied to a variety of membranes for the isolation of specific proteins.

Original languageEnglish
Pages (from-to)209-212
Number of pages4
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume823
Issue number2
DOIs
Publication statusPublished - 2005 Sep 5
Externally publishedYes

Keywords

  • Affinity chromatography
  • Divalent cations
  • Heparin binding proteins
  • Membrane proteins

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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