Abstract
Human 5-lipoxygenase requires ATP as a stimulatory factor. At the two preferred concentrations of the free Ca2+, 0.02 μM with a resting cell and 20 μM with a stimulated cell, Scatchard analysis revealed that 5-lipoxygenase has one affinity ATP binding site with aK d of 4.6 μM at the low Ca2+ concentration but has two affinity ATP binding sites with a higherK d of 4.4 μM and a lowerK d of 14.5 μM at the high Ca2+ concentration. In contrast, in a Tween 20 reaction system, 5-lipoxygenase had similar activation coefficients for ATP at both Ca2+ concentrations; these were 12.7 μM at the low Ca2+ concentration and 12.0 μM at the high Ca2+ concentration. These results showed that 5-lipoxygenase has an ATP binding site and suggest that self-association of 5-lipoxygenase in 20 μM Ca2+ may affect ATP binding affinity as measured by Scatchard analysis.
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Abbreviations
- α-S-ATP:
-
adenosine 5′-(α-thio) triphosphate
- γ-S-ATP:
-
adenosine 5′-(γ-thio) triphosphate
- 5-HPETE:
-
5S-hydroperoxy-6-trans-8,11,14-cis-icosatetraenoic acid
- LTA4 :
-
5,6-trans-oxido-7,9-trans-11,14-cis-icosatetraenoic acid
- LTB4 :
-
5S,12R-dihydroxy-6,14-cis-8,10-trans-icosatetraenoic acid
- NTA:
-
nitrilotriacetic acid
- Tween 20:
-
polyoxyethylene sorbitanmonolaurate
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Noguchi, M., Miyano, M. & Matsumoto, T. Physicochemical characterization of ATP binding to human 5-lipoxygenase. Lipids 31, 367–371 (1996). https://doi.org/10.1007/BF02522921
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DOI: https://doi.org/10.1007/BF02522921