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 335166
Title Distinct conformational stability and functional activity of four highly homologous endonuclease colicins
Author(s) Bremer, E.T.J. van den; Keeble, A.H.; Jiskoot, W.; Spelbrink, R.E.J.; Maier, C.S.; Hoek, A. van; Visser, A.J.W.G.; James, R.; Moore, G.R.; Kleanthous, C.; Heck, A.J.R.
Source Protein Science 13 (2004)5. - ISSN 0961-8368 - p. 1391 - 1401.
DOI https://doi.org/10.1110/ps.03508204
Department(s) Biophysics
Biochemistry
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2004
Keyword(s) ionization mass-spectrometry - protein-protein interactions - electrospray-ionization - immunity protein - transition-metals - crystal-structure - escherichia-coli - nuclease domain - e9 - complexes
Abstract The family of conserved colicin DNases E2, E7, E8, and E9 are microbial toxins that kill bacteria through random degradation of the chromosomal DNA. In the present work, we compare side by side the conformational stabilities of these four highly homologous colicin DNases. Our results indicate that the apo-forms of these colicins are at room temperature and neutral pH in a dynamic conformational equilibrium between at least two quite distinct conformers. We show that the thermal stabilities of the apo-proteins differ by up to 20degreesC. The observed differences correlate with the observed conformational behavior, that is, the tendency of the protein to form either an open, less stable or closed, more stable conformation in solution, as deduced by both tryptophan accessibility studies and electrospray ionization mass spectrometry. Given these surprising structural differences, we next probed the catalytic activity of the four DNases and also observed a significant variation in relative activities. However, no unequivocal link between the activity of the protein and its thermal and structural stability could easily be made. The observed differences in conformational and functional properties of the four colicin DNases are surprising given that they are a closely related ( greater than or equal to65% identity) family of enzymes containing a highly conserved (betabetaalpha-Me) active site motif. The different behavior of the apo-enzymes must therefore most likely depend on more subtle changes in amino acid sequences, most likely in the exosite region (residues 72-98) that is required for specific high-affinity binding of the cognate immunity protein.
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