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 335167
Title Ligand-induced changes in the conformational dynamics of a bacterial cytotoxic endonuclease
Author(s) Bremer, E.T.J. van den; Keeble, A.H.; Visser, A.J.W.G.; Hoek, A. van; Kleanthous, C.; Heck, A.J.R.; Jiskoot, W.
Source Biochemistry 43 (2004)14. - ISSN 0006-2960 - p. 4347 - 4355.
Department(s) Biochemistry
Publication type Refereed Article in a scientific journal
Publication year 2004
Keyword(s) protein-protein interactions - single-tryptophan mutants - n-h motif - colicin e9 - transition-metals - immunity proteins - escherichia-coli - dnase domain - active-site - complex
Abstract Knowledge about the conformational dynamics of a protein is key to understanding its biochemical and biophysical properties. In the present work we investigated the dynamic properties of the enzymatic domain of DNase colicins via time-resolved fluorescence and anisotropy decay analysis in combination with steady-state acrylamide quenching experiments. The dynamic properties of the apoenzyme were compared to those of the E9 DNase ligated to the transition metal ion Zn2+ and the natural inhibitor Im9. We further investigated the contributions of each of the two tryptophans within the E9 DNase (Trp22 and Trp58) using two single-tryptophan mutants (E9 W22F and E9 W58F). Wild-type E9 DNase, E9 W22F, and E9 W58F, as well as Im9, showed multiple lifetime decays. The time-resolved and steady-state fluorescence results indicated that complexation of E9 DNase with Zn2+ induces compaction of the E9 DNase structure, accompanied by immobilization of Trp22 along with a reduced solvent accessibility for both tryptophans. Im9 binding resulted in immobilization of Trp22 along with a decrease in the longest lifetime component. In contrast, Trp58 experienced less restriction on complexation of E9 DNase with Im9 and showed an increase in the longest lifetime component. Furthermore, the results point out that the Im9-induced changes in the conformational dynamics of E9 DNase are predominant and occur independently of the Zn2+-induced conformational effects.
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