The Three Isoforms of Hepcidin in Human Serum and Their Processing Determined by Liquid Chromatography-tandem Mass Spectrometry (LC-tandem MS)

Overview

Journal Int J Hematol

Specialty Hematology

Date 2015 Oct 15

PMID 26462810

Citations 9

Authors

Lynda Addo

Katsuya Ikuta

Hiroki Tanaka

Yasumichi Toki

Mayumi Hatayama

Masayo Yamamoto

Satoshi Ito

Motohiro Shindo

Yusuke Sasaki

Yasushi Shimonaka

Mikihiro Fujiya

Yutaka Kohgo

Affiliations

Soon will be listed here.

Abstract

Hepcidin, the iron regulatory hormone, has three isoforms; -20, -22 and -25. While hepcidin-25 has been studied extensively, the physiological significance of other isoforms remains poorly understood. Using a quantitative method based on liquid chromatography-tandem mass spectrometry (LC-tandem MS) developed by our group, we quantified hepcidin isoforms in human serum to elucidate their characteristics, and investigated the role of hepatocytes in isoform processing. Hepcidin isoforms in serum obtained from 40 healthy volunteers were quantified. Synthetic hepcidin peptides were added to healthy serum, and to HepG2 culture media, and hepcidin isoform concentrations determined. All three hepcidin isoforms were detected in human serum; however, hepcidin-25 concentrations were highest. The three hepcidin isoforms showed a strong positive correlation with each other and with serum ferritin. Additionally, while hepcidin-20 was strongly correlated with serum creatinine, the other isoforms were not. Hepcidin-20 and -25 levels were also increased in chronic kidney disease (CKD) serum. Hepcidin-22 rapidly degraded into hepcidin-20, whereas hepcidin-25 remained relatively stable. Finally, hepcidin-22 degradation into hepcidin-20 was accelerated in the presence of HepG2. This method has enabled us to reveal fundamental characteristics of the three hepcidin isoforms in serum and may be a powerful tool for quantifying hepcidin isoform expression and processing.

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