| Autor: |
Hulla, Franz W., Höckel, Michael, Risi, Sergio, Dose, Klaus |
| Předmět: |
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| Zdroj: |
European Journal of Biochemistry; Aug76 Part 2, Vol. 67 Issue 2, p469-476, 8p |
| Abstrakt: |
A chemically reactive ATP analogue. 6-[(3-carboxy-4-nitrophenyl)thio]-9-β-D-ribofuranosylpurine 5′-triphosphate (Nbs6ITP) has been synthesized. It has the ability to form stable thioether bonds between the 6-position of the purine ring and aliphatic mercapto groups. The nucleotide moiety of the reagent has been covalently bound to agarose, via iminobispropylamine and N-acetyl-homocysteine as spacer with the purpose of producing an affinity chromatography material. The affinity matrix binds solubilized F1 ATPase from a crude extract of Micrococcus sp. membranes. Afterwards the enzyme can be selectively eluted from the column at a defined ATP concentration. This method is superior to the conventional purification with respect to speed and convenience of the preparation. The affinity chromatography leads in a one-step process to the same purity of enzyme, substituting several steps of the conventional method. In addition, the affinity matrix was used for binding studies. Although the presence of Mg2+ ions is a prerequisite for the hydrolysis of nucleoside 5′-triphosphates. evidence is presented indicating that the binding of the nucleoside triphosphates to highly purified F1 ATPase from Micrococcus sp. appears not to be influenced by Mg2+ ion concentrations so far examined. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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