Free-solution interaction assay of carbonic anhydrase to its inhibitors using back-scattering interferometry.
Back-scattering interferometry (BSI) is a label-free, free-solution, small-volume technique used for characterizing binding interactions, which is also relevant to a growing number of biosensing applications including drug discovery. Here, we use BSI to characterize the interaction of carbonic anhydrase enzyme II with five well-known carbonic anhydrase enzyme II inhibitors (± sulpiride, sulfanilamide, benzene sulfonamide, dansylamide, and acetazolamide) in the presence of DMSO. Dissociation constants calculated for each interaction were consistent with literature values previously obtained using surface plasmon resonance and fluorescence-based competition assays. Results demonstrate the potential of BSI as a drug-screening tool which is fully compatible with DMSO and does not require immobilization or labeling, therefore allowing binding interactions to be characterized in the native state. BSI has the potential for reducing labor costs, sample consumption, and assay time while providing enhanced reliability over existing techniques.
Department of Chemistry and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, USA.
Ereny F
EF
Morcos
Amanda
A
Kussrow
Carolyn
C
Enders
Darryl
D
Bornhop
D000086:Acetazolamide / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000378:metabolism; D003619:Dansyl Compounds / Q000737:chemistry / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D007368:Interferometry / Q000379:methods*; D011485:Protein Binding; D012542:Scattering, Radiation; D013424:Sulfanilamides / Q000737:chemistry / Q000378:metabolism
D002257:Carbonic Anhydrase Inhibitors; D003619:Dansyl Compounds; D013424:Sulfanilamides; C001026:5-dimethylaminonaphthalene-1-sulfonamide; D002256:Carbonic Anhydrases; D000086:Acetazolamide; D004121:Dimethyl Sulfoxide