title: An efficient strategy to enhance binding affinity and specificity of a known isozyme inhibitor.
abstract: The binding profile of a known inhibitor, benzenesulfonamide, against a family of carbonic anhydrase isozymes was efficiently enhanced via high-throughput screening of customized combinatorial one-bead-one-compound peptide libraries modified with the inhibitor molecule. The screening of the conjugate libraries recognized subtle variations in the microenvironments of the target enzyme and thus facilitated the identification of short peptide sequences that bind selectively to a close proximity of the active site. The identified peptide portions contributed significantly to the overall binding of the conjugate peptides with greatly enhanced affinity as well as improved specificity towards the target isozyme. The interactions between the inhibitors and the isozymes were validated by surface plasmon resonance (SPR), pull-down assay and enzymatic activity measurement. This high-throughput approach proved useful and efficient to enhance the binding profile of known inhibitors and may apply to developing effective inhibitors for a wide range of isozyme families.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Joo-Eun
authors.initials: JE
authors.lastname: Jee
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Jaehong
authors.initials: J
authors.lastname: Lim
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Yong Siang
authors.initials: YS
authors.lastname: Ong
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Jessica
authors.initials: J
authors.lastname: Oon
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Liqian
authors.initials: L
authors.lastname: Gao
authors.affiliation: Gordon Center for Medical Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 55 Fruit Street, White 427, Boston, MA 02114, USA. hchoi12@mgh.harvard.edu.
authors.forename: Hak Soo
authors.initials: HS
authors.lastname: Choi
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Su Seong
authors.initials: SS
authors.lastname: Lee
mesh_terms: D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D062105:Molecular Docking Simulation; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D011485:Protein Binding; D013449:Sulfonamides / Q000737:chemistry* / Q000494:pharmacology*; D000096926:Benzenesulfonamides
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D002256:Carbonic Anhydrases; D007527:Isoenzymes; D019151:Peptide Library; D010455:Peptides; D013449:Sulfonamides; D000096926:Benzenesulfonamides