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

doi:10.1016/j.jchromb.2005.06.025

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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

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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 Mg(2+) in the first and increasing concentrations of Ca(2+) 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.

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Amino Acid Sequence Carrier Proteins Cations, Divalent Cell Fractionation Chromatography, Affinity Electrophoresis, Polyacrylamide Gel HL-60 Cells Heparin Humans Membrane Proteins Mitochondrial Proteins Reproducibility of Results Sequence Analysis, Protein Journal Article

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Tsukimi Iida, Masaharu Kamo, Nobuyuki Uozumi, Takashi Inui, Katsuyuki Imai. 2005-09-05. 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.. https://doi.org/10.1016/j.jchromb.2005.06.025

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