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Studies on the interaction of firefly luciferase with triazine dyes

Authors
Journal
Journal of Chromatography A
0021-9673
Publisher
Elsevier
Publication Date
Volume
355
Identifiers
DOI: 10.1016/s0021-9673(01)97316-9
Disciplines
  • Biology

Abstract

Abstract The interaction of firefly luciferase with several triazine dyes has been investigated. Of those tested, the anthraquinone dye, Procion blue MX-R, displayed relatively high affinity [dissociation constant ( K D)  5μ M] and irreversibly inactivated the enzyme in a time dependent fashion. The substrates, MgATP (ATP is adenosine triphosphate) and luciferin, protected the enzyme against inactivation with Procion blue MX-R and following quantitative inactivation, incorporation of 1 mol dye per mol enzyme [molecular weight ( M r)  50 000] was observed. It is suggested that Procion blue MX-R binding is active site-directed and that the dye binding traverses both MgATP and luciferin binding sites. These data are exploited to evolve an improved purification protocol for firefly luciferase.

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