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
| Autor: | Takashi Inui, Nobuyuki Uozumi, Katsuyuki Imai, Masaharu Kamo, Tsukimi Iida |
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| Rok vydání: | 2004 |
| Předmět: |
Cations
Divalent Clinical Biochemistry HL-60 Cells Cell Fractionation Biochemistry DNA-binding protein Chromatography Affinity Analytical Chemistry Divalent Mitochondrial Proteins chemistry.chemical_compound Affinity chromatography Sequence Analysis Protein Humans Amino Acid Sequence Polyacrylamide gel electrophoresis chemistry.chemical_classification Octyl glucoside Chromatography Chemistry Elution Heparin Membrane Proteins Reproducibility of Results Cell Biology General Medicine Membrane Membrane protein Electrophoresis Polyacrylamide Gel Carrier Proteins |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 823(2) |
| ISSN: | 1570-0232 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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