Metal-binding molecules in the organs of Mus musculus by size-exclusion chromatography coupled with UV spectroscopy and ICP-MS

Abstract

Mus musculus mice have been investigated for the total elements content in different organs (lung, liver, spleen, kidney, brain, testicle, heart and muscle) and molecular mass distribution patterns of Mn, Ni, Cu, Zn, As, Pb, Cr, Fe, Co, Se and Cd. Some differences have been found in the organs studied, with especially relevant being the Cu-containing fraction present only in the brain and the As-containing one in the liver. Other differences related to the abundance of the metallospecies have also been found. The present paper is the first step in the study of the "metallome" of this inbred laboratory species from which the genome is completely known. This organism could be used as a model in future studies focused on wild mice and the analytical approach developed could be applied to wild mice to find markers of environmental pollution. [figure: see text] The present paper is the first step in the study of the "metallome" of the inbred laboratory specie Mus musculus from which the genome is completely known. Some interesting differences have been found in the extracts from the organs that are discussed along the text.

Liquid chromatography-inductively coupled plasma-based metallomic approaches to probe health-relevant interactions between xenobiotics and mammalian organisms

“…In addition, all agents that exert chelating properties and may therefore form complexes with metal ions that are weakly-bound to proteins during sample preparation procedures, such as EDTA (often used as an anti-coagulant) and the reducing agent dithiothreitol (DTT; is frequently used in sample preparation procedures in conventional proteomics protocols) must be strictly avoided. 7,40,70,71 Instead of EDTA one should use heparin as the anti-coagulant when plasma has to be prepared from blood. 62 Similarly, it is critical to replace DTT by reduced glutathione (GSH) and to employ a GSH concentration in the homogenization buffer that closely resembles the concentration of GSH in the cytosol of the cells of interest in order not to adversely affect chemical equilibria.…”

Mercury and selenium binding biomolecules in terrestrial mammals (Cervus elaphus and Sus scrofa) from a mercury exposed area

“…To perform the metal binding molecule analysis in biological tissues, an extraction step is necessary to liberate the molecules from the matrix, and transfer it into the separation and detection system. This step is critical, as the most important goal is to keep the metal-biomolecule integrity, avoiding the rupture of metal-biomolecule bonds or the exchange of metals [22,23]. Therefore, it is necessary to optimize variables in the stage of sample preparation, such as the extraction agent used, the concentration of extracting agent, extraction volume, homogenization time and pH of the medium, to obtain a maximum extraction efficiency whilst keeping the integrity of the metal-biomolecule entity.…”

Metallomics in environmental and health related research: Current status and perspectives

“…Limit reports have achieved the global analysis of metalloproteome in specific biological samples, for example investigation of metalloproteome of Pyrococcus furiosus, Escherichia coli, and Sulfolobus solfataricus, which clearly implied microbial metalloproteomes were largely unknown [46]. Most of works either monitored the metal contained proteins which however not characterized the proteins identity, or focused on particular proteins [45,[88][89][90]. Uncharacterized insights into the behaviors and toxicity of metals were still provided.…”