Distribution of Co, Cu, Fe, Mn, Se, Zn, and Cd among cytosolic proteins of different molecular masses in gills of European chub (Squalius cephalus L.)
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The distribution of essential elements Co, Cu, Fe, Mn, Se, and Zn, and nonessential element Cd among cytosolic proteins of different molecular masses in the gills of European chub (Squalius cephalus) sampled in the moderately contaminated Sutla River in September of 2009, was studied after the protein separation by size exclusion high-performance liquid chromatography (SEC-HPLC), and the metal determination in the obtained fractions by high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS). The aims of the study were to characterize the distribution profiles of metals within different protein categories in gills in the conditions of low metal exposure in the river water, and to compare them with the previously published hepatic profiles. The distribution profiles of analyzed metals were mainly characterized with several peaks. However, some observations could be emphasized: both Cu and Cd were eluted near metallothionein elution time; elution time of one of Co peaks could be associated with Co-containing compound cobalamin; increasing cytosolic Fe concentrations resulted in possible Fe binding to storage protein ferritin; both Mn and Zn had poorly resolved peaks covering wide ranges of molecular masses and indicating their binding to various proteins; both Zn and Se increased in protein fractions of molecular masses <5 kDa following their concentration increase in the gill cytosol; expected clear metallothionein peak was not observed for Zn. Comparison of gill profiles with previously published hepatic profiles revealed similar and in case of some elements (e.g., Co, Fe, Mn, and Se) almost identical distributions in both organs regarding elution times. On the contrary, heights of obtained peaks were different, indicating possible metal binding to the same proteins in the gills and liver, but in different proportions. The results obtained in this study can be used as a basis for comparison in monitoring studies, for identification of changes that would occur after exposure of chub to increased metal concentrations.
KeywordsCytosolic proteins European chub Gills HR ICP-MS Metals SEC-HPLC
The financial support by the Ministry of Science, Education and Sport of the Republic of Croatia (project no. 098-0982934-2721 ) is acknowledged. This study was carried out as a part of the Monitoring of freshwater fishery in 2009—Group D—Fishing area Sava River–Sutla River, funded by the Ministry of Agriculture, Fisheries and Rural Development of the Republic of Croatia. Special thanks are due to Branislav Španović, Dr. Damir Valić, Dr. Damir Kapetanović, Dr. Irena Vardić Smrzlić, Dr. Božidar Kurtović, and Dr. Emin Teskeredžić for collaboration on the common fish sampling; to Željka Strižak, B. Sc. for the assistance in the field work and metal analyses; as well as to Dr. Nevenka Mikac from Laboratory for Inorganic Environmental Geochemistry for the opportunity to use HR ICP-MS .
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