Publication

Models to determine the kinetic mechanisms of ion-coupled transporters

Lolkema, J. S. & Slotboom, D. J., 10-Jan-2019, In : The Journal of General Physiology. 151, 3, p. 369-380 11 p., jgp.201812055.

Research output: Contribution to journalArticleAcademicpeer-review

With high-resolution structures available for many ion-coupled (secondary active) transporters, a major challenge for the field is to determine how coupling is accomplished. Knowledge of the kinetic mechanism of the transport reaction, which defines the binding order of substrate and co-ions, together with the sequence with which all relevant states are visited by the transporter, will help to reveal this coupling mechanism. Here, we derived general mathematical models that can be used to analyze data from steady-state transport measurements and show how kinetic mechanisms can be derived. The models describe how the apparent maximal rate of substrate transport depends on the co-ion concentration, and vice versa, in different mechanisms. Similarly, they describe how the apparent affinity for the transported substrate is affected by the co-ion concentration and vice versa. Analyses of maximal rates and affinities permit deduction of the number of co-ions that bind before, together with, and after the substrate. Hill analysis is less informative, but in some mechanisms, it can reveal the total number of co-ions transported with the substrate. However, prior knowledge of the number of co-ions from other experimental approaches is preferred when deriving kinetic mechanisms, because the models are generally overparameterized. The models we present have wide applicability for the study of ion-coupled transporters.

Original languageEnglish
Article numberjgp.201812055
Pages (from-to)369-380
Number of pages11
JournalThe Journal of General Physiology
Volume151
Issue number3
Early online date10-Jan-2019
Publication statusPublished - 10-Jan-2019

    Keywords

  • kinetic mechanisms, ion-coupled transporters

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