title: Structure and binding properties of a cameloid nanobody raised against KDM5B.
abstract: The histone demethylase KDM5B is considered to be a promising target for anticancer therapy. Single-chain antibodies from llama (nanobodies) have been raised to aid in crystallization and structure determination of this enzyme. The antigen-binding properties of 15 of these nanobodies have been characterized. The crystal structure of one of these (NB17) has been determined to a resolution of 1.85 Å. NB17 crystallizes in space group P4322 with six molecules in the asymmetric unit. The six molecules in the asymmetric unit pack as an entity with approximate D3 symmetry with interactions mediated by the CDR loops, which could act as a crystallization nucleus. NB17 does not bind to monomeric KDM5B residues 1-820, but is found to bind to aggregates formed after incubation at 310 K.
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Anders
authors.initials: A
authors.lastname: Wiuf
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Line Hyltoft
authors.initials: LH
authors.lastname: Kristensen
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Ole
authors.initials: O
authors.lastname: Kristensen
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Dorosz
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Jonas
authors.initials: J
authors.lastname: Jensen
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark.
authors.forename: Michael
authors.initials: M
authors.lastname: Gajhede
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002162:Camelus; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000737:chemistry* / Q000378:metabolism*; D013696:Temperature
chemical_list: D009687:Nuclear Proteins; D012097:Repressor Proteins; D061905:Single-Domain Antibodies; D056484:Jumonji Domain-Containing Histone Demethylases; C119611:KDM5B protein, human
keywords: KDM5B; nanobody