title: Functional characterization of fluorescent hepcidin.
abstract: Hepcidin is a peptide hormone that regulates homeostasis in iron metabolism. It binds to the sole known cellular iron exporter ferroportin (Fpn), triggers its internalization, and thereby modulates the efflux of iron from cells. This functional property has been adopted in this study to assess the bioactivity and potency of a range of novel fluorescent hepcidin analogues. Hepcidin was selectively labeled with 6-carboxyfluorescein (CF) and 6-carboxytetramethylrhodamine (TMR) using Fmoc solid phase peptide chemistry. Internalization of Fpn by hepcidin was assessed by high-content microscopic analysis. Both K18- and M21K-labeled hepcidin with TMR and CF exhibited measurable potency when tested in cultured MDCK and T47D cells expressing human ferroportin. The bioactivity of the labeled hepcidin varies with the type of fluorophore and site of attachment of the fluorophores on the hepcidin molecule.
authors.affiliation: Chemical Biology Group, Institute of Pharmaceutical Sciences, and ‡Nutrition and Diabetes Research Group, King's College London , Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, U.K.
authors.forename: Franz
authors.initials: F
authors.lastname: Dürrenberger
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Abbate
authors.forename: Yongmin
authors.initials: Y
authors.lastname: Ma
authors.forename: Maria C
authors.initials: MC
authors.lastname: Arno
authors.forename: Dareen
authors.initials: D
authors.lastname: Jaiash
authors.forename: Archna
authors.initials: A
authors.lastname: Parmar
authors.forename: Victoria
authors.initials: V
authors.lastname: Marshall
authors.forename: Gladys O
authors.initials: GO
authors.lastname: Latunde-Dada
authors.forename: Tina
authors.initials: T
authors.lastname: Zimmermann
authors.forename: David
authors.initials: D
authors.lastname: Senn
authors.forename: Patrick
authors.initials: P
authors.lastname: Altermatt
authors.forename: Vania
authors.initials: V
authors.lastname: Manolova
authors.forename: Robert C
authors.initials: RC
authors.lastname: Hider
authors.forename: Sukhvinder S
authors.initials: SS
authors.lastname: Bansal
mesh_terms: D000818:Animals; D027682:Cation Transport Proteins / Q000378:metabolism; D002460:Cell Line; D004285:Dogs; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D064451:Hepcidins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D017510:Protein Folding; D012235:Rhodamines / Q000737:chemistry; D000098742:Ferroportin
chemical_list: C106141:6-carboxytetramethylrhodamine; D027682:Cation Transport Proteins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D064451:Hepcidins; D012235:Rhodamines; D000098742:Ferroportin; C024098:6-carboxyfluorescein