Iron-binding biomolecules in the soluble hepatic fraction of the northern pike (Esox lucius): two-dimensional chromatographic separation with mass spectrometry detection

Dragun, Zrinka; Kiralj, Zoran; Ivanković, Dušica; Bilić, Branka; Kazazić, Saša; Kazazić, Snježana (2024) Iron-binding biomolecules in the soluble hepatic fraction of the northern pike (Esox lucius): two-dimensional chromatographic separation with mass spectrometry detection. Analytical and Bioanalytical Chemistry, 416 . pp. 5097-5109. ISSN 1618-2650

	PDF - Accepted Version - article Restricted to Closed Access until 24 July 2025. Download (1MB) | Request a personal copy from author
	PDF - Accepted Version - research data Restricted to Closed Access until 24 July 2025. Download (1MB) | Request a personal copy from author

Abstract

Iron plays vital roles in important biological processes in fish, but can be toxic in high concentrations. The information on metalloproteins that participate in maintenance of Fe homeostasis in an esocid fish, the northern pike, as an important freshwater bioindicator species, are rather scarce. The aim of this study was to identify main cytosolic constituents that sequester Fe in the northern pike liver. The method applied consisted of two-dimensional HPLC separation of Fe-binding biomolecules, based on anion-exchange followed by size exclusion fractionation. Apparent molecular masses of two main Fe-metalloproteins isolated by this procedure were 360 kDa and 50 kDa, with the former having more acidic pI, and indicated presence of ferritin and haemoglobin, respectively. MALDI-TOF-MS provided confirmation of ferritin subunit with a m/z peak at 20.65 kDa, and haemoglobin with spectra containing main m/z peak at 16.1 kDa, and smaller peaks at 32.1, 48.2, and 7.95 kDa (single charged Hb-monomer, dimer, and trimer, and double-charged monomer, respectively). LC MS/MS with subsequent MASCOT database search, confirmed the presence of Hb-subunits and pointed to close relation between esocid and salmonid fishes. Further efforts should be directed towards optimization of the conditions for metalloprotein analysis by mass spectrometry, to extend the knowledge on intracellular metal-handling mechanisms.

Item Type:	Article
Uncontrolled Keywords:	ferritin; fish; haemoglobin; HPLC; liver; MALDI-TOF-MS
Subjects:	NATURAL SCIENCES > Chemistry > Analytic Chemistry NATURAL SCIENCES > Chemistry > Applied Chemistry NATURAL SCIENCES > Biology > Biochemistry and Molecular Biology NATURAL SCIENCES > Interdisciplinary Natural Sciences NATURAL SCIENCES > Interdisciplinary Natural Sciences > Environmental Science
Divisions:	Division for Marine and Enviromental Research
Projects:	Project title Project leader Project code Project type Biomolekule koje vežu metale i zdravstveni poremećaji kod slatkovodnih organizama izloženih industrijskom otpadu-METABIOM Zrinka Dragun IP-2019-04-2636 HRZZ
Depositing User:	Zrinka Dragun
Date Deposited:	16 Jan 2025 15:08
URI:	http://fulir.irb.hr/id/eprint/9476
DOI:	10.1007/s00216-024-05446-y

Actions (login required)

View Item