Improved Accuracy of Low Affinity Protein–Ligand Equilibrium Dissociation Constants Directly Determined by Electrospray Ionization Mass Spectrometry
| Autor: | Fabienne Saab, Martine Cadene, Lucie Jaquillard, Françoise Schoentgen |
|---|---|
| Přispěvatelé: | Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de minéralogie et de physique des milieux condensés (IMPMC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Institut de Physique du Globe de Paris (IPG Paris)-Centre National de la Recherche Scientifique (CNRS), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Université Pierre et Marie Curie - Paris 6 (UPMC)-IPG PARIS-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2012 |
| Předmět: |
Models
Molecular Spectrometry Mass Electrospray Ionization Electrospray ionization Kinetics Analytical chemistry Ligands 010402 general chemistry Mass spectrometry Kinetic energy MESH: Spectrometry Mass Electrospray Ionization 01 natural sciences Dissociation (chemistry) Structural Biology MESH: Ligands Animals Humans MESH: Protein Binding MESH: Animals MESH: Proteins [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Spectroscopy MESH: Gases MESH: Humans MESH: Kinetics Nucleotides Ligand Chemistry 010401 analytical chemistry MESH: Chickens Proteins MESH: Nucleotides 0104 chemical sciences Dissociation constant MESH: Cattle Cattle Gases Chickens MESH: Models Molecular Protein Binding Protein ligand |
| Zdroj: | Journal of The American Society for Mass Spectrometry Journal of The American Society for Mass Spectrometry, 2012, 23 (5), pp.908-22. ⟨10.1007/s13361-011-0305-7⟩ Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), 2012, 23 (5), pp.908-22. ⟨10.1007/s13361-011-0305-7⟩ |
| ISSN: | 1044-0305 1879-1123 |
| DOI: | 10.1007/s13361-011-0305-7 |
| Popis: | International audience; There is continued interest in the determination by ESI-MS of equilibrium dissociation constants (K(D)) that accurately reflect the affinity of a protein-ligand complex in solution. Issues in the measurement of K(D) are compounded in the case of low affinity complexes. Here we present a K(D) measurement method and corresponding mathematical model dealing with both gas-phase dissociation (GPD) and aggregation. To this end, a rational mathematical correction of GPD (f(sat)) is combined with the development of an experimental protocol to deal with gas-phase aggregation. A guide to apply the method to noncovalent protein-ligand systems according to their kinetic behavior is provided. The approach is validated by comparing the K(D) values determined by this method with in-solution K(D) literature values. The influence of the type of molecular interactions and instrumental setup on f(sat) is examined as a first step towards a fine dissection of factors affecting GPD. The method can be reliably applied to a wide array of low affinity systems without the need for a reference ligand or protein. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink