title: Surface plasmon resonance biosensor assay for the analysis of small-molecule inhibitor binding to human and parasitic phosphodiesterases.
abstract: In the past decade, surface plasmon resonance (SPR) biosensor-based technology has been exploited more and more to characterize the interaction between drug targets and small-molecule modulators. Here, we report the successful application of SPR methodology for the analysis of small-molecule binding to two therapeutically relevant cAMP phosphodiesterases (PDEs), Trypanosoma brucei PDEB1 which is implicated in African sleeping sickness and human PDE4D which is implicated in a plethora of disease conditions including inflammatory pulmonary disorders such as asthma, chronic obstructive pulmonary disease and central nervous system (CNS) disorders. A protocol combining the use of directed capture using His-tagged PDE_CDs with covalent attachment to the SPR surface was developed. This methodology allows the determination of the binding kinetics of small-molecule PDE inhibitors and also allows testing their specificity for the two PDEs. The SPR-based assay could serve as a technology platform for the development of highly specific and high-affinity PDE inhibitors, accelerating drug discovery processes.
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands. Electronic address: m.siderius@vu.nl.
authors.forename: Marco
authors.initials: M
authors.lastname: Siderius
authors.affiliation: IOTA Pharmaceuticals, St. John's Innovation Centre, Cowley Road, Cambridge CB4 0WS, United Kingdom.
authors.forename: Anitha
authors.initials: A
authors.lastname: Shanmugham
authors.affiliation: IOTA Pharmaceuticals, St. John's Innovation Centre, Cowley Road, Cambridge CB4 0WS, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: England
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Tiffany
authors.initials: T
authors.lastname: van der Meer
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Jan Paul
authors.initials: JP
authors.lastname: Bebelman
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Antoni R
authors.initials: AR
authors.lastname: Blaazer
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands; IOTA Pharmaceuticals, St. John's Innovation Centre, Cowley Road, Cambridge CB4 0WS, United Kingdom.
authors.forename: Iwan J P
authors.initials: IJ
authors.lastname: de Esch
authors.affiliation: Amsterdam Institute of Molecules, Medicines and Systems (AIMMS), Division of Medicinal Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands; IOTA Pharmaceuticals, St. John's Innovation Centre, Cowley Road, Cambridge CB4 0WS, United Kingdom.
authors.forename: Rob
authors.initials: R
authors.lastname: Leurs
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D010726:Phosphodiesterase Inhibitors / Q000032:analysis* / Q000737:chemistry*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000737:chemistry* / Q000378:metabolism; D054852:Small Molecule Libraries / Q000032:analysis* / Q000737:chemistry*; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance
chemical_list: D010726:Phosphodiesterase Inhibitors; D015800:Protozoan Proteins; D054852:Small Molecule Libraries; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; C517376:PDE4D protein, human; C581006:PDEB1 protein, Trypanosoma brucei
keywords: African sleeping disease; Phosphodiesterase; Small molecules; Surface plasmon resonance; Trypanosoma brucei