Journal article


How does Mg2+ affect the binding of anhydrotetracycline in the TetR protein?


Publication Details
Author(s): Leypold CF, Marian DT, Roman C, Schneider S, Schubert P, Scholz O, Hillen W, Clark T, Lanig H
Publisher: Royal Society of Chemistry
Publication year: 2004
Volume: 3
Journal issue: 1
Pages range: 109-119
ISSN: 1474-905X

Abstract

The binding of anhydrotetracycline (atc) in wild-type TetR( D), TetR( B), and four single tryptophan mutants of TetR( B) was investigated by UV/vis absorption, steady-state, and time-resolved fluorescence spectroscopy. From absorption titration experiments with Mg2+, we conclude that binding of one [atc-Mg](+) complex in the homodimer causes changes in the protein conformation around the second binding pocket. In the presence or absence of Mg2+, several different groups of atc-protein arrangements must exist, each with a characteristic atc fluorescence decay time. Taking into account the results of molecular dynamics ( MD) simulations, we propose as one possible origin for such a differentiation the extent of hydrogen bonding between atc and the surrounding amino acids. Binding of Mg2+ should change the arrangement of the surrounding amino acids such that some of the excited atc molecules do not undergo the relaxation processes typical for free atc. The MD simulations also show that the pattern of intra- and intermolecular hydrogen bonding in the two monomeric units is not correlated, thereby leading to different fluorescence kinetics for atc in the two monomeric units. Furthermore, it is suggested that hydrogen bonding between Arg104 and O10 of anhydrotetracycline could regulate the relaxation processes of excited anhydrotetracycline.

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