title: The elution of certain protein affinity tags with millimolar concentrations of diclofenac.
abstract: Diclofenac (2-[(2, 6-dichlorophenyl)amino] benzeneacetic acid) is a sparingly soluble, nonsteroidal anti-inflammatory drug therapeutically acting at low micromolar concentrations. In pH range from 8 to 11, its aqueous solubility can be increased up to 200 times by the presence of counter ions such as sodium. Our protein interaction studies revealed that a millimolar concentration of sodium diclofenac is able to elute glutathione S-transferase (GST), cellulose binding protein (CBD), and maltose binding protein (MBP) but not histidine-tagged or PDZ-tagged proteins from their affinity resins. The elution efficiency of diclofenac is comparable with the eluting agents normally used at similar concentrations. Native gel electrophoresis of sodium diclofenac-treated proteins showed that the interaction is non-covalent and non-denaturing. These results suggest that sodium diclofenac, in addition to its pharmaceutical applications, can also be exploited as a lead for the development of new proteomics reagents.
authors.affiliation: Laboratory of Neurobiology, Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovakia.
authors.forename: Martina
authors.initials: M
authors.lastname: Baliova
authors.affiliation: Laboratory of Neurobiology, Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovakia.
authors.forename: Anna
authors.initials: A
authors.lastname: Juhasova
authors.affiliation: Laboratory of Neurobiology, Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovakia. Electronic address: Frantisek.Jursky@savba.sk.
authors.forename: Frantisek
authors.initials: F
authors.lastname: Jursky
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D004008:Diclofenac / Q000737:chemistry* / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry* / Q000378:metabolism; D058619:Maltose-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D015203:Reproducibility of Results; D012995:Solubility
chemical_list: D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D004008:Diclofenac; D005982:Glutathione Transferase
keywords: CBD; Cellulose binding protein; Diclofenac; GST; Glutathione S-transferase; MBP; Maltose binding protein; PDZ (PSD95/Discs large/ZO-1)