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Mechanism of Indium(III) Exchange between NTA and Transferrin: A Kinetic Approach

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    Mechanism of Indium(III) Exchange between NTA and Transferrin: A Kinetic Approach
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    Dipartimento di Chimica e Chimica Industriale, University of Pisa, Via Risorgimento 35, 56100 Pisa, Italy
    * To whom correspondence should be addressed. Telephone: +39-050-2219259. Fax: +39-050-2219260. E-mail: [email protected]
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    The Journal of Physical Chemistry B

    Cite this: J. Phys. Chem. B 2008, 112, 38, 12168–12173
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    https://doi.org/10.1021/jp8045033
    Published August 30, 2008
    Copyright © 2008 American Chemical Society
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    Abstract

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    The equilibria and kinetics for the process of In3+ exchange between nitrilotriacetic acid (NTA) and bovine serum transferrin (T) have been investigated in aqueous solution containing sodium bicarbonate. The metal exchange equilibria have been measured by difference ultraviolet spectroscopy at 25 °C, pH = 7.4, and I = 0.2 M (NaClO4). The acid dissociation constants of NTA and the binding constants of In(III) to NTA have also been measured. Kinetic experiments revealed that the process of In3+ uptake by transferrin from [In(NTA)2]3- is biphasic, the fast phase being completed in a few seconds, the slow phase lasting for hours. The fast phase has been investigated by the stopped-flow method and results in monoexponential kinetics. It involves rapid interaction of the 1:1 complex ML (M = In, L = NTA) with TB (T = transferrin, B = CO32−) to give a quaternary intermediate MLTB which then evolves to an “open” MTB* ternary complex complex with expulsion of L. In turn, this complex interconverts to a “closed”, more stable, form MTB. Neither the prevailing complex M2L nor the TB2 form of transferrin are directly involved in the exchange process but act as metal and protein reservoirs. The pH dependence of the reaction has been also investigated. The slow phase has not been investigated in detail; it takes several hours to go to the completeness, its slowness being ascribed to metal redistribution between the C-site and N-site of the protein, and/or metal release from polynuclear In(III) species.

    Copyright © 2008 American Chemical Society

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    Supporting Information

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    Figures showing potentiometric titration curves of NTA and In−NTA, species distribution plots for NTA, indium−NTA complexes, and indium−NTA−transferrin systems, and the difference spectra of transferrin in the presence of different amounts of [In(NTA)2]3−, text giving the mathematical derivation of eqs 3, 4, and 5 and a scheme showing the reactions used in the derivation of eq 5. This material is available free of charge via the Internet at http://pubs.acs.org.

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    Cited By

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    The Journal of Physical Chemistry B

    Cite this: J. Phys. Chem. B 2008, 112, 38, 12168–12173
    Click to copy citationCitation copied!
    https://doi.org/10.1021/jp8045033
    Published August 30, 2008
    Copyright © 2008 American Chemical Society
    Request reuse permissions

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