title: The high-molecular-weight kininogen domain 5 is an intrinsically unstructured protein and its interaction with ferritin is metal mediated.
abstract: High-molecular-weight kininogen domain 5 (HK5) is an angiogenic modulator that is capable of inhibiting endothelial cell proliferation, migration, adhesion, and tube formation. Ferritin can bind to a histidine-glycine-lysine-rich region within HK5 and block its antiangiogenic effects. However, the molecular intricacies of this interaction are not well understood. Analysis of the structure of HK5 using circular dichroism and nuclear magnetic resonance [(1) H, (15) N]-heteronuclear single quantum coherence determined that HK5 is an intrinsically unstructured protein, consistent with secondary structure predictions. Equilibrium binding studies using fluorescence anisotropy were used to study the interaction between ferritin and HK5. The interaction between the two proteins is mediated by metal ions such as Co(2+) , Cd(2+) , and Fe(2+) . This metal-mediated interaction works independently of the loaded ferrihydrite core of ferritin and is demonstrated to be a surface interaction. Ferritin H and L bind to HK5 with similar affinity in the presence of metals. The ferritin interaction with HK5 is the first biological function shown to occur on the surface of ferritin using its surface-bound metals.
authors.affiliation: Center for Structural Biology, Wake Forest School of Medicine, Winston-Salem, North Carolina; Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, North Carolina.
authors.forename: Annissa J
authors.initials: AJ
authors.lastname: Huhn
authors.forename: Derek
authors.initials: D
authors.lastname: Parsonage
authors.forename: David A
authors.initials: DA
authors.lastname: Horita
authors.forename: Frank M
authors.initials: FM
authors.lastname: Torti
authors.forename: Suzy V
authors.initials: SV
authors.lastname: Torti
authors.forename: Thomas
authors.initials: T
authors.lastname: Hollis
mesh_terms: D005293:Ferritins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019679:Kininogen, High-Molecular-Weight / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019216:Metals, Heavy / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular
chemical_list: D064267:Intrinsically Disordered Proteins; D019679:Kininogen, High-Molecular-Weight; D019216:Metals, Heavy; D005293:Ferritins
keywords: angiogenesis; ferritin; intrinsically unstructured protein; kininogen; metal ions; protein interaction