Mass spectrometry analysis of Fe-binding cytosolic biomolecules from the liver of the northern pike (Esox lucius) after two-dimensional chromatographic separation

Kiralj, Zoran; Dragun, Zrinka; Ivanković, Dušica; Bilić, Branka; Kazazić, Saša; Kazazić, Snježana Mass spectrometry analysis of Fe-binding cytosolic biomolecules from the liver of the northern pike (Esox lucius) after two-dimensional chromatographic separation. In: 15th Mass Spectrometry School in Biotechnology and Medicine (2 July 2023 - 8 July 2023) Dubrovnik. (Unpublished)

Preview

Image (JPEG) - Published Version - poster Download (177kB) | Preview

Abstract

The size-exclusion (SEC) fractionation of Fe-binding biomolecules from the northern pike hepatic cytosol, using HPLC coupled to ICP-MS, revealed that numerous proteins participate in Fe-cellular metabolism. Since cytosolic Co/Mo-binding biomolecules have similar masses to those binding Fe, two-dimensional separation (anion-exchange (AEX) and SEC) was applied prior to mass spectrometry analyses. The Fe-fraction with AEX-maximum at 9.2 min (SEC-estimated molecular mass: 53.3 kDa) was revelaed by MALDI-TOF-MS to contain peaks at 16.05, 32.12, and 48.19 kDa, indicating hemoglobin monomer, dimer and trimer. LC-MS/MS investigation with database search confirmed presence of hemoglobin -subunit (comparable to subunits from Esox lucius and Oncorhyncus nerka). The Fe-fraction with AEX-maximum at 10.6 min (SEC-estimated molecular mass: 360 kDa) was, by comparison with apoferritin standard, hypothesized to contain ferritin, the primary Fe-storage protein. The combination of mass spectrometry analyses with multidimensional chromatographic separation proves to be promising tool for gaining better insight in cellular behaviour of metals.

Item Type:	Unpublished conference/workshop items or lecture materials
Uncontrolled Keywords:	mass spectrometry; liquid chromatography; ICP-MS; northern pike; MALDI-TOF-MS; LC-MS/MS
Subjects:	INTERDISCIPLINARY AREAS OF KNOWLEDGE
Divisions:	Division for Marine and Enviromental Research Division of Physical Chemistry
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:	Zoran Kiralj
Date Deposited:	01 Oct 2024 09:09
URI:	http://fulir.irb.hr/id/eprint/9092

Actions (login required)

View Item