title: Steric hindrance as a basis for structure-based design of selective inhibitors of protein-tyrosine phosphatases.
abstract: Utilizing structure-based design, we have previously demonstrated that it is possible to obtain selective inhibitors of protein-tyrosine phosphatase 1B (PTP1B). A basic nitrogen was introduced into a general PTP inhibitor to form a salt bridge to Asp48 in PTP1B and simultaneously cause repulsion in PTPs containing an asparagine in the equivalent position [Iversen, L. F., et al. (2000) J. Biol. Chem. 275, 10300-10307]. Further, we have recently demonstrated that Gly259 in PTP1B forms the bottom of a gateway that allows easy access to the active site for a broad range of substrates, while bulky residues in the same position in other PTPs cause steric hindrance and reduced substrate recognition capacity [Peters, G. H., et al. (2000) J. Biol. Chem. 275, 18201-18209]. The current study was undertaken to investigate the feasibility of structure-based design, utilizing these differences in accessibility to the active site among various PTPs. We show that a general, low-molecular weight PTP inhibitor can be developed into a highly selective inhibitor for PTP1B and TC-PTP by introducing a substituent, which is designed to address the region around residues 258 and 259. Detailed enzyme kinetic analysis with a set of wild-type and mutant PTPs, X-ray protein crystallography, and molecular modeling studies confirmed that selectivity for PTP1B and TC-PTP was achieved due to steric hindrance imposed by bulky position 259 residues in other PTPs.
authors.affiliation: Protein Chemistry and Signal Transduction, Novo Nordisk, DK-2880 Bagsvaerd, Denmark. lfiv@novonordisk.com
authors.forename: L F
authors.initials: LF
authors.lastname: Iversen
authors.forename: H S
authors.initials: HS
authors.lastname: Andersen
authors.forename: K B
authors.initials: KB
authors.lastname: Møller
authors.forename: O H
authors.initials: OH
authors.lastname: Olsen
authors.forename: G H
authors.initials: GH
authors.lastname: Peters
authors.forename: S
authors.initials: S
authors.lastname: Branner
authors.forename: S B
authors.initials: SB
authors.lastname: Mortensen
authors.forename: T K
authors.initials: TK
authors.lastname: Hansen
authors.forename: J
authors.initials: J
authors.lastname: Lau
authors.forename: Y
authors.initials: Y
authors.lastname: Ge
authors.forename: D D
authors.initials: DD
authors.lastname: Holsworth
authors.forename: M J
authors.initials: MJ
authors.lastname: Newman
authors.forename: N P
authors.initials: NP
authors.lastname: Hundahl Møller
mesh_terms: D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D017433:Protein Structure, Secondary; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; C516607:PTPN1 protein, human; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases