Disturbed natural balance as a result of significant pollution pressure by mining activities has ... more Disturbed natural balance as a result of significant pollution pressure by mining activities has been already reported for the major waterways in the Republic of Macedonia. An exceeded metal concentration in the aquatic environment has the potential to induce serious harm in wild organisms, to impair their health, condition, reproduction or their habitats. The implementation of the EU Water Framework Directive establishes that the ecological status of the aquatic ecosystems must be determined by biological quality elements, including fish, and has to be supported by hydro-morphological and physico-chemical parameters. Therefore, the aim of this study was to obtain basic data for the effect of mining activities on fish, using Vardar chub (Squalius vardarensis) as a bioindicator organism, and using biomarkers that are relatively simple and non-specific, but indicative of cumulative organism-level changes. For achieving this goal, 158 chub were captured with electro fishing methods fro...
Metals play crucial physiological roles, but they can also cause irreparable toxic effects throug... more Metals play crucial physiological roles, but they can also cause irreparable toxic effects through binding to important cellular biomolecules in aquatic organisms. The aim of this study was to determine the exact molecular masses and to identify several selected metal-binding biomolecules in hepatic and gill cytosols of Vardar chub (Squalius vardarensis Karaman, 1928). Methods applied for the achievement of this goal were SEC-AEC-HPLC for two-dimensional separation of cytosolic biomolecules, HR ICP-MS for metal measurements, and mass spectrometry (MALDI-TOF-MS and LC-MS/MS) for biomolecule mass determination and identification. The analyzed biomolecules included: Fe-binding biomolecules, which were identified as hemoglobin subunit b in the liver (molecular masses of B15 kDa), and hemoglobin subunits a and b in the gills (molecular masses of B11 kDa, B13 kDa and B15 kDa); heat-stable Cdbinding biomolecules, which were identified as MT isoforms MT-I and MT-II (molecular mass of B6.0 kDa in both liver and gills, and an additional 4.9 kDa isoform in the gills); and heat-stable Mo-binding biomolecules of molecular masses equal to 3.3 kDa (in the gills) and 8.5 kDa (in the liver). An important finding of this study was the obvious presence of different isoforms of the same biomolecules in the liver and gills. This was, among others, manifested through the absence of Zn binding to MTs only in the gills, indicating that the same type of biomolecule can be responsible for different functions in different organs. Thus, for better understanding of metal behaviour in aquatic organisms, it is crucial to identify cellular metal-binding biomolecules and their functions.
The distribution of essential elements Co, Cu, Fe,
Mn, Se, and Zn, and nonessential element Cd am... more 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 nearmetallothionein 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.
Association of selected essential (Co, Cu, Fe,
Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trac... more Association of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements with cytosolic proteins of different molecular masses was described for the liver of European chub (Squalius cephalus) from weakly contaminated Sutla River in Croatia. The principal aim was to establish basic trace element distributions among protein fractions characteristic for the fish living in the conditions of low metal exposure in the water. The fractionation of chub hepatic cytosols was carried out by size exclusion high performance liquid chromatography (SE-HPLC; Superdex™ 200 10/300 GL column), and measurements were performed by high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Elution profiles of essential elements were mostly characterized by broad peaks covering wide range of molecular masses, as a sign of incorporation of essential elements in various proteins within hepatic cytosol. Exceptions were Cu and Fe, with elution profiles characterized by sharp, narrow peaks indicating their probable association with specific proteins, metallothionein (MT), and ferritin, respectively. The main feature of the elution profile of nonessential metal Cd was also single sharp, narrowpeak, coinciding with MT elution time, and indicating almost complete Cd detoxification by MT under the conditions of weak metal exposure in the water (dissolved Cd concentration ≤0.3 μg L−1). Contrary, nonessential metal Pb was observed to bind to wide spectrum of proteins, mostly of medium molecular masses (30– 100 kDa), after exposure to dissolved Pb concentration of ~1 μg L−1. The obtained information within this study presents the starting point for identification and characterization of specific metal/metalloid-binding proteins in chub hepatic cytosol, which could be further used as markers of metal/ metalloid exposure or effect on fish.
• Metallomics approach in monitoring effects of metal pollution on Vardar chub • Use of SEC-HPLC/... more • Metallomics approach in monitoring effects of metal pollution on Vardar chub • Use of SEC-HPLC/HR ICP-MS for determination of metal distributions within cytosol • Distribution profiles of Cd, Co, Cu, Fe, Mn, Mo, Se and Zn in chub liver and gills • Changes in the distribution profiles due to increased metal exposure level in water • Comparison of metal profiles in Vardar and European chub, two related fish species G R A P H I C A L A B S T R A C T a b s t r a c t Many bioindicators have not yet been well characterized regarding their tendency to bind trace elements by different cytosolic biomolecules in response to trace element exposure. Accordingly, our principal aim was to define the cytosolic distributions of Cd, Co, Cu, Fe, Mn, Mo, Se, and Zn among the biomolecules of different molecular masses in liver and gills of Vardar chub (Squalius vardarensis Karaman), a representative fish species of Macedo-nian rivers, and to determine distribution changes which occur as a consequence of increased exposure to specific trace elements. Additionally, we aimed to confirm the presence of heat-stable biomolecules in chub hepatic and gill cytosols. Distribution profiles were obtained by separation of cytosols and heat-treated cytosols using size-exclusion high performance-liquid chromatography, and by offline determination of trace element concentrations using high resolution inductively coupled plasma-mass spectrometry. Distribution profiles of trace elements were mainly characterized by several peaks encompassing different ranges of molecular masses, as a sign of incorporation of trace elements in various biomolecules within hepatic and gill cytosols. Especially interesting finding was probable binding of Fe to ferritin, which was especially pronounced in the liver, as a sign of important liver function in Fe storage. Furthermore, association with heat-stable proteins, metallothioneins (MT), was indicated for Cd, Cu, and Zn in the hepatic cytosol, as well as for Cd in the gill cytosol, whereas a sign of Zn-MT association was not observed in the gills. The presence of Mo-and Se-binding heat-stable compounds of very low molecular masses (b 10 kDa) in the cytosol was determined for both liver and the gills. Trace elements under j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / s c i t o t e n v all studied conditions were found associated to the same biomolecules, and only their proportions associated to specific cytosolic compounds have changed as a consequence of their increased bioaccumulation in the liver and gills of Vardar chub.
Disturbed natural balance as a result of significant pollution pressure by mining activities has ... more Disturbed natural balance as a result of significant pollution pressure by mining activities has been already reported for the major waterways in the Republic of Macedonia. An exceeded metal concentration in the aquatic environment has the potential to induce serious harm in wild organisms, to impair their health, condition, reproduction or their habitats. The implementation of the EU Water Framework Directive establishes that the ecological status of the aquatic ecosystems must be determined by biological quality elements, including fish, and has to be supported by hydro-morphological and physico-chemical parameters. Therefore, the aim of this study was to obtain basic data for the effect of mining activities on fish, using Vardar chub (Squalius vardarensis) as a bioindicator organism, and using biomarkers that are relatively simple and non-specific, but indicative of cumulative organism-level changes. For achieving this goal, 158 chub were captured with electro fishing methods fro...
Metals play crucial physiological roles, but they can also cause irreparable toxic effects throug... more Metals play crucial physiological roles, but they can also cause irreparable toxic effects through binding to important cellular biomolecules in aquatic organisms. The aim of this study was to determine the exact molecular masses and to identify several selected metal-binding biomolecules in hepatic and gill cytosols of Vardar chub (Squalius vardarensis Karaman, 1928). Methods applied for the achievement of this goal were SEC-AEC-HPLC for two-dimensional separation of cytosolic biomolecules, HR ICP-MS for metal measurements, and mass spectrometry (MALDI-TOF-MS and LC-MS/MS) for biomolecule mass determination and identification. The analyzed biomolecules included: Fe-binding biomolecules, which were identified as hemoglobin subunit b in the liver (molecular masses of B15 kDa), and hemoglobin subunits a and b in the gills (molecular masses of B11 kDa, B13 kDa and B15 kDa); heat-stable Cdbinding biomolecules, which were identified as MT isoforms MT-I and MT-II (molecular mass of B6.0 kDa in both liver and gills, and an additional 4.9 kDa isoform in the gills); and heat-stable Mo-binding biomolecules of molecular masses equal to 3.3 kDa (in the gills) and 8.5 kDa (in the liver). An important finding of this study was the obvious presence of different isoforms of the same biomolecules in the liver and gills. This was, among others, manifested through the absence of Zn binding to MTs only in the gills, indicating that the same type of biomolecule can be responsible for different functions in different organs. Thus, for better understanding of metal behaviour in aquatic organisms, it is crucial to identify cellular metal-binding biomolecules and their functions.
The distribution of essential elements Co, Cu, Fe,
Mn, Se, and Zn, and nonessential element Cd am... more 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 nearmetallothionein 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.
Association of selected essential (Co, Cu, Fe,
Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trac... more Association of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements with cytosolic proteins of different molecular masses was described for the liver of European chub (Squalius cephalus) from weakly contaminated Sutla River in Croatia. The principal aim was to establish basic trace element distributions among protein fractions characteristic for the fish living in the conditions of low metal exposure in the water. The fractionation of chub hepatic cytosols was carried out by size exclusion high performance liquid chromatography (SE-HPLC; Superdex™ 200 10/300 GL column), and measurements were performed by high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Elution profiles of essential elements were mostly characterized by broad peaks covering wide range of molecular masses, as a sign of incorporation of essential elements in various proteins within hepatic cytosol. Exceptions were Cu and Fe, with elution profiles characterized by sharp, narrow peaks indicating their probable association with specific proteins, metallothionein (MT), and ferritin, respectively. The main feature of the elution profile of nonessential metal Cd was also single sharp, narrowpeak, coinciding with MT elution time, and indicating almost complete Cd detoxification by MT under the conditions of weak metal exposure in the water (dissolved Cd concentration ≤0.3 μg L−1). Contrary, nonessential metal Pb was observed to bind to wide spectrum of proteins, mostly of medium molecular masses (30– 100 kDa), after exposure to dissolved Pb concentration of ~1 μg L−1. The obtained information within this study presents the starting point for identification and characterization of specific metal/metalloid-binding proteins in chub hepatic cytosol, which could be further used as markers of metal/ metalloid exposure or effect on fish.
• Metallomics approach in monitoring effects of metal pollution on Vardar chub • Use of SEC-HPLC/... more • Metallomics approach in monitoring effects of metal pollution on Vardar chub • Use of SEC-HPLC/HR ICP-MS for determination of metal distributions within cytosol • Distribution profiles of Cd, Co, Cu, Fe, Mn, Mo, Se and Zn in chub liver and gills • Changes in the distribution profiles due to increased metal exposure level in water • Comparison of metal profiles in Vardar and European chub, two related fish species G R A P H I C A L A B S T R A C T a b s t r a c t Many bioindicators have not yet been well characterized regarding their tendency to bind trace elements by different cytosolic biomolecules in response to trace element exposure. Accordingly, our principal aim was to define the cytosolic distributions of Cd, Co, Cu, Fe, Mn, Mo, Se, and Zn among the biomolecules of different molecular masses in liver and gills of Vardar chub (Squalius vardarensis Karaman), a representative fish species of Macedo-nian rivers, and to determine distribution changes which occur as a consequence of increased exposure to specific trace elements. Additionally, we aimed to confirm the presence of heat-stable biomolecules in chub hepatic and gill cytosols. Distribution profiles were obtained by separation of cytosols and heat-treated cytosols using size-exclusion high performance-liquid chromatography, and by offline determination of trace element concentrations using high resolution inductively coupled plasma-mass spectrometry. Distribution profiles of trace elements were mainly characterized by several peaks encompassing different ranges of molecular masses, as a sign of incorporation of trace elements in various biomolecules within hepatic and gill cytosols. Especially interesting finding was probable binding of Fe to ferritin, which was especially pronounced in the liver, as a sign of important liver function in Fe storage. Furthermore, association with heat-stable proteins, metallothioneins (MT), was indicated for Cd, Cu, and Zn in the hepatic cytosol, as well as for Cd in the gill cytosol, whereas a sign of Zn-MT association was not observed in the gills. The presence of Mo-and Se-binding heat-stable compounds of very low molecular masses (b 10 kDa) in the cytosol was determined for both liver and the gills. Trace elements under j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / s c i t o t e n v all studied conditions were found associated to the same biomolecules, and only their proportions associated to specific cytosolic compounds have changed as a consequence of their increased bioaccumulation in the liver and gills of Vardar chub.
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Papers by Nesrete Krasnići
to important cellular biomolecules in aquatic organisms. The aim of this study was to determine the
exact molecular masses and to identify several selected metal-binding biomolecules in hepatic and gill
cytosols of Vardar chub (Squalius vardarensis Karaman, 1928). Methods applied for the achievement of
this goal were SEC-AEC-HPLC for two-dimensional separation of cytosolic biomolecules, HR ICP-MS
for metal measurements, and mass spectrometry (MALDI-TOF-MS and LC-MS/MS) for biomolecule mass
determination and identification. The analyzed biomolecules included: Fe-binding biomolecules, which
were identified as hemoglobin subunit b in the liver (molecular masses of B15 kDa), and hemoglobin
subunits a and b in the gills (molecular masses of B11 kDa, B13 kDa and B15 kDa); heat-stable Cdbinding
biomolecules, which were identified as MT isoforms MT-I and MT-II (molecular mass of B6.0
kDa in both liver and gills, and an additional 4.9 kDa isoform in the gills); and heat-stable Mo-binding
biomolecules of molecular masses equal to 3.3 kDa (in the gills) and 8.5 kDa (in the liver). An important
finding of this study was the obvious presence of different isoforms of the same biomolecules in the
liver and gills. This was, among others, manifested through the absence of Zn binding to MTs only in the
gills, indicating that the same type of biomolecule can be responsible for different functions in different
organs. Thus, for better understanding of metal behaviour in aquatic organisms, it is crucial to identify
cellular metal-binding biomolecules and their functions.
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 nearmetallothionein
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.
Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace
elements with cytosolic proteins of different molecular
masses was described for the liver of European chub
(Squalius cephalus) from weakly contaminated Sutla
River in Croatia. The principal aim was to establish
basic trace element distributions among protein fractions
characteristic for the fish living in the conditions of low
metal exposure in the water. The fractionation of chub
hepatic cytosols was carried out by size exclusion high
performance liquid chromatography (SE-HPLC;
Superdex™ 200 10/300 GL column), and measurements
were performed by high resolution inductively coupled
plasma mass spectrometry (HR ICP-MS). Elution profiles
of essential elements were mostly characterized by
broad peaks covering wide range of molecular masses,
as a sign of incorporation of essential elements in various
proteins within hepatic cytosol. Exceptions were
Cu and Fe, with elution profiles characterized by sharp,
narrow peaks indicating their probable association with
specific proteins, metallothionein (MT), and ferritin, respectively.
The main feature of the elution profile of
nonessential metal Cd was also single sharp, narrowpeak, coinciding with MT elution time, and indicating
almost complete Cd detoxification by MT under the
conditions of weak metal exposure in the water (dissolved
Cd concentration ≤0.3 μg L−1). Contrary, nonessential
metal Pb was observed to bind to wide spectrum
of proteins, mostly of medium molecular masses (30–
100 kDa), after exposure to dissolved Pb concentration of
~1 μg L−1. The obtained information within this study
presents the starting point for identification and characterization
of specific metal/metalloid-binding proteins in chub hepatic
cytosol, which could be further used as markers of metal/
metalloid exposure or effect on fish.
to important cellular biomolecules in aquatic organisms. The aim of this study was to determine the
exact molecular masses and to identify several selected metal-binding biomolecules in hepatic and gill
cytosols of Vardar chub (Squalius vardarensis Karaman, 1928). Methods applied for the achievement of
this goal were SEC-AEC-HPLC for two-dimensional separation of cytosolic biomolecules, HR ICP-MS
for metal measurements, and mass spectrometry (MALDI-TOF-MS and LC-MS/MS) for biomolecule mass
determination and identification. The analyzed biomolecules included: Fe-binding biomolecules, which
were identified as hemoglobin subunit b in the liver (molecular masses of B15 kDa), and hemoglobin
subunits a and b in the gills (molecular masses of B11 kDa, B13 kDa and B15 kDa); heat-stable Cdbinding
biomolecules, which were identified as MT isoforms MT-I and MT-II (molecular mass of B6.0
kDa in both liver and gills, and an additional 4.9 kDa isoform in the gills); and heat-stable Mo-binding
biomolecules of molecular masses equal to 3.3 kDa (in the gills) and 8.5 kDa (in the liver). An important
finding of this study was the obvious presence of different isoforms of the same biomolecules in the
liver and gills. This was, among others, manifested through the absence of Zn binding to MTs only in the
gills, indicating that the same type of biomolecule can be responsible for different functions in different
organs. Thus, for better understanding of metal behaviour in aquatic organisms, it is crucial to identify
cellular metal-binding biomolecules and their functions.
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 nearmetallothionein
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.
Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace
elements with cytosolic proteins of different molecular
masses was described for the liver of European chub
(Squalius cephalus) from weakly contaminated Sutla
River in Croatia. The principal aim was to establish
basic trace element distributions among protein fractions
characteristic for the fish living in the conditions of low
metal exposure in the water. The fractionation of chub
hepatic cytosols was carried out by size exclusion high
performance liquid chromatography (SE-HPLC;
Superdex™ 200 10/300 GL column), and measurements
were performed by high resolution inductively coupled
plasma mass spectrometry (HR ICP-MS). Elution profiles
of essential elements were mostly characterized by
broad peaks covering wide range of molecular masses,
as a sign of incorporation of essential elements in various
proteins within hepatic cytosol. Exceptions were
Cu and Fe, with elution profiles characterized by sharp,
narrow peaks indicating their probable association with
specific proteins, metallothionein (MT), and ferritin, respectively.
The main feature of the elution profile of
nonessential metal Cd was also single sharp, narrowpeak, coinciding with MT elution time, and indicating
almost complete Cd detoxification by MT under the
conditions of weak metal exposure in the water (dissolved
Cd concentration ≤0.3 μg L−1). Contrary, nonessential
metal Pb was observed to bind to wide spectrum
of proteins, mostly of medium molecular masses (30–
100 kDa), after exposure to dissolved Pb concentration of
~1 μg L−1. The obtained information within this study
presents the starting point for identification and characterization
of specific metal/metalloid-binding proteins in chub hepatic
cytosol, which could be further used as markers of metal/
metalloid exposure or effect on fish.