Functional Characterization of Fluorescent Hepcidin

Franz Durrenberger; Vincenzo Abbate; Yongmin Ma; Maria C Arno; Dareen  Jaiash; Archna Parmar; Victoria Marshall; Gladys Latunde-Dada; Tina Zimmerman; David Senn; Patrick Altermatt; Vania  Manolova; Robert Hider; Sukhi Bansal

doi:10.1021/bc400121x

Functional Characterization of Fluorescent Hepcidin

Franz Durrenberger, Vincenzo Abbate, Yongmin Ma, Maria C Arno, Dareen Jaiash, Archna Parmar, Victoria Marshall, Gladys Latunde-Dada, Tina Zimmerman, David Senn, Patrick Altermatt, Vania Manolova, Robert Hider, Sukhi Bansal

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7 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1527–1532
Journal	Bioconjugate Chemistry
Volume	24
Issue number	9
Early online date	26 Jul 2013
DOIs	https://doi.org/10.1021/bc400121x
Publication status	Published - 18 Sept 2013

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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.",

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doi = "10.1021/bc400121x",

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TY - JOUR

T1 - Functional Characterization of Fluorescent Hepcidin

AU - Durrenberger, Franz

AU - Abbate, Vincenzo

AU - Ma, Yongmin

AU - Arno, Maria C

AU - Jaiash, Dareen

AU - Parmar, Archna

AU - Marshall, Victoria

AU - Latunde-Dada, Gladys

AU - Zimmerman, Tina

AU - Senn, David

AU - Altermatt, Patrick

AU - Manolova, Vania

AU - Hider, Robert

AU - Bansal, Sukhi

PY - 2013/9/18

Y1 - 2013/9/18

N2 - 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.

AB - 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.

U2 - 10.1021/bc400121x

DO - 10.1021/bc400121x

M3 - Article

SN - 1043-1802

VL - 24

SP - 1527

EP - 1532

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 9

ER -

Functional Characterization of Fluorescent Hepcidin

Abstract

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