Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 386050
Title The intrinsic fluorescence of apo-obelin and apo-aequorin and use of its quenching to characterize coelenterazine binding.
Author(s) Eremeeva, E.; Markova, S.V.; Westphal, A.H.; Visser, A.J.W.G.; Berkel, W.J.H. van; Vysotski, S.
Source FEBS Letters 583 (2009)12. - ISSN 0014-5793 - p. 1939 - 1944.
Department(s) Biochemistry
Publication type Refereed Article in a scientific journal
Publication year 2009
Keyword(s) crystal-structure - ca2+-regulated photoproteins - violet bioluminescence - angstrom resolution - recombinant obelin - w92f obelin - calcium - regeneration - apoaequorin - expression
Abstract The intrinsic fluorescence of two apo-photoproteins has been characterized and its concentration-dependent quenching by coelenterazine has been for the first time applied to determine the apparent dissociation constants for coelenterazine binding with apo-aequorin (1.2±0.12µM) and apo-obelin (0.2±0.04µM). Stopped-flow measurements of fluorescence quenching showed that coelenterazine binding is a millisecond-scale process, in contrast to the formation of an active photoprotein complex taking several hours. This finding evidently shows that the rate-limiting step of active photoprotein formation is the conversion of coelenterazine into its 2-hydroperoxy derivative
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