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 427684
Title Distant residues mediate picomolar binding affinity of a protein cofactor
Author(s) Bollen, Y.J.M.; Westphal, A.H.; Lindhoud, S.; Berkel, W.J.H. van; Mierlo, C.P.M. van
Source Nature Communications 3 (2012). - ISSN 2041-1723
DOI http://dx.doi.org/10.1038/ncomms2010
Department(s) Biochemistry
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) azotobacter-vinelandii apoflavodoxin - hydrogen-exchange - ligand-binding - oxidized flavodoxin - energy landscape - nmr relaxation - dynamics - kinetics - thermodynamics - topology
Abstract Numerous proteins require cofactors to be active. Computer simulations suggest that cooperative interaction networks achieve optimal cofactor binding. There is a need for the experimental identification of the residues crucial for stabilizing these networks and thus for cofactor binding. Here we investigate the electron transporter flavodoxin, which contains flavin mononucleotide as non-covalently bound cofactor. We show that after binding flavin mononucleotide with nanomolar affinity, the protein relaxes extremely slowly (time constant ~5 days) to an energetically more favourable state with picomolar-binding affinity. Rare small-scale openings of this state are revealed through H/D exchange of N(3)H of flavin. We find that H/D exchange can pinpoint amino acids that cause tight cofactor binding. These hitherto unknown residues are dispersed throughout the structure, and many are located distantly from the flavin and seem irrelevant to flavodoxin's function. Quantification of the thermodynamics of ligand binding is important for understanding, engineering, designing and evolving ligand-binding proteins
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