title: Structural analysis of lipocalin-type prostaglandin D synthase complexed with biliverdin by small-angle X-ray scattering and multi-dimensional NMR.
abstract: Lipocalin-type prostaglandin D synthase (L-PGDS) acts as both a PGD(2) synthase and an extracellular transporter for small lipophilic molecules. From a series of biochemical studies, it has been found that L-PGDS has an ability to bind a variety of lipophilic ligands such as biliverdin, bilirubin and retinoids in vitro. Therefore, we considered that it is necessary to clarify the molecular structure of L-PGDS upon binding ligand in order to understand the physiological relevance of L-PGDS as a transporter protein. We investigated a molecular structure of L-PGDS/biliverdin complex by small-angle X-ray scattering (SAXS) and multi-dimensional NMR measurements, and characterized the binding mechanism in detail. SAXS measurements revealed that L-PGDS has a globular shape and becomes compact by 1.3A in radius of gyration on binding biliverdin. NMR experiments revealed that L-PGDS possessed an eight-stranded antiparallel beta-barrel forming a central cavity. Upon the titration with biliverdin, some cross-peaks for residues surrounding the cavity and EF-loop and H2-helix above the beta-barrel shifted, and the intensity of other cross-peaks decreased with signal broadenings in (1)H-(15)N heteronuclear single quantum coherence spectra. These results demonstrate that L-PGDS holds biliverdin within the beta-barrel, and the conformation of the loop regions above the beta-barrel changes upon binding biliverdin. Through such a conformational change, the whole molecule of L-PGDS becomes compact.
authors.affiliation: Laboratory of Protein Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Miyamoto
authors.forename: Shigenori
authors.initials: S
authors.lastname: Nishimura
authors.forename: Katsuaki
authors.initials: K
authors.lastname: Inoue
authors.forename: Shigeru
authors.initials: S
authors.lastname: Shimamoto
authors.forename: Takuya
authors.initials: T
authors.lastname: Yoshida
authors.forename: Ayano
authors.initials: A
authors.lastname: Fukuhara
authors.forename: Mao
authors.initials: M
authors.lastname: Yamada
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Urade
authors.forename: Naoto
authors.initials: N
authors.lastname: Yagi
authors.forename: Tadayasu
authors.initials: T
authors.lastname: Ohkubo
authors.forename: Takashi
authors.initials: T
authors.lastname: Inui
mesh_terms: D000818:Animals; D001664:Biliverdine / Q000737:chemistry* / Q000378:metabolism; D005453:Fluorescence; D019746:Intramolecular Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D054834:Lipocalins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008956:Models, Chemical; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: D054834:Lipocalins; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase; D001664:Biliverdine