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      Influence of the Lipid Phase Removal on the Quantification of Cu, Fe and Zn Bound to Proteins from Golden Flaxseed (<italic>Linum usitatissimum L</italic>.)

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          Abstract

          <p>The analysis of metals bound to proteins requires careful sample preparation. Experimental procedures involved in this area should be developed aiming to retain the metal species bound to the protein structure. Some pre-fractionation steps are commonly used and in many cases it is necessary to disrupt the lipid-protein interaction by performing lipid removal before protein extraction. This paper presents a systematic investigation on the influence of using hexane, chloroform and methanol neat or in sequence to remove the lipid fraction in golden flaxseed (<italic>Linum usitatissimum L.</italic>), before extracting the protein phase using Tris-HCl buffer solution for analysis of Cu, Fe and Zn bound to proteins. The results showed that the organic solvent used has a strong influence on the determination of these metals. With hexane, the total protein content and Cu, Fe and Zn concentrations in Tris-HCl phase were higher than those found using methanol:chloroform (1:2 v/v).</p>

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          Most cited references25

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          Orientações sobre Validação de Métodos de Ensaios Químicos

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            A simple set-and-mix assay for screening of protein kinase inhibitors in cell lysates.

            Here we developed a simple set-and-mix assay to perform high-throughput screening of protein kinase A (PKA) inhibitors from the LOPAC 1280 compound library. This assay is based on the color change of gold nanoparticles on aggregation induced by a cationic substrate peptide as coagulant. In spite of the simplicity of this assay system, this assay can be applied to drug screening based on cellular kinases. We successfully found several highly active inhibitors, including compounds that have not been reported before.
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              Quim. Nova

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                jbchs
                Journal of the Brazilian Chemical Society
                J. Braz. Chem. Soc.
                Sociedade Brasileira de Química (São Paulo )
                1678-4790
                August 2015
                : 26
                : 8
                : 1718-1724
                Affiliations
                [1 ] Universidade Federal de Minas Gerais Brazil
                Article
                S0103-50532015000801718
                10.5935/0103-5053.20150146
                ea935c78-d51d-495d-afb8-330110239453

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0103-5053&lng=en
                Categories
                CHEMISTRY, MULTIDISCIPLINARY

                General chemistry
                metal,protein,sample preparation,lipid phase removal,organic solvent
                General chemistry
                metal, protein, sample preparation, lipid phase removal, organic solvent

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