Selective removal of copper from complex biological media with an agarose-immobilized high-affinity PSP ligand

IF 2.7 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

JBIC Journal of Biological Inorganic Chemistry Pub Date : 2024-07-27 DOI:10.1007/s00775-024-02065-x

Arielle Nabatilan, M. Thomas Morgan, Sara Netzer, Christoph J. Fahrni

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Abstract

The elucidation of metal-dependent biological processes requires selective reagents for manipulating metal ion levels within biological solutions such as growth media or cell lysates. To this end, we immobilized a phosphine sulfide-stabilized phosphine (PSP) ligand on agarose to create a resin for the selective removal of copper from chemically complex biological media through simple filtration or centrifugation. Comprised of a conformationally preorganized phenylene-bridged backbone, the PSP-ligand binds Cu(I) with a 1:1 stoichiometry and exhibits a pH-independent Cu(I) dissociation constant in the low zeptomolar range. Neither Zn(II), Fe(II), nor Mn(II) interact with the ligand at millimolar concentrations, thus offering a much-improved selectivity towards copper over other commonly employed solid-supported chelators such as Chelex 100. As revealed by X-ray fluorescence elemental analysis, the immobilized chelator effectively removes copper from cell culture growth media and cell lysate isolated from mouse fibroblasts. In addition to preparing copper-depleted media or cell lysates for biological studies, PSP-immobilized ligands might prove equally useful for applications in radiochemistry, materials science, and environmental science.

Graphical abstract

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利用琼脂糖固定化高亲和性 PSP 配体从复杂生物介质中选择性去除铜

阐明依赖金属的生物过程需要选择性试剂，以控制生物溶液（如生长培养基或细胞裂解液）中的金属离子水平。为此，我们将硫化膦-稳定膦（PSP）配体固定在琼脂糖上，制成了一种树脂，可通过简单过滤或离心从化学性质复杂的生物介质中选择性地去除铜。PSP 配体由构象预组织的苯基桥接骨架组成，能以 1:1 的配比结合铜(I)，并在低七摩尔范围内表现出与 pH 值无关的铜(I)解离常数。在毫摩尔浓度下，Zn(II)、Fe(II)和Mn(II)都不会与配体发生作用，因此与其他常用的固体支持螯合剂（如Chelex 100）相比，它对铜的选择性大大提高。X 射线荧光元素分析表明，固定化螯合剂能有效去除细胞培养生长介质和从小鼠成纤维细胞中分离出来的细胞裂解物中的铜。除了制备用于生物研究的贫铜培养基或细胞裂解物外，PSP 固定化配体在放射化学、材料科学和环境科学领域的应用也同样有用。

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来源期刊

JBIC Journal of Biological Inorganic Chemistry 化学-生化与分子生物学

CiteScore

5.90

自引率

3.30%

发文量

审稿时长

3 months

期刊介绍： Biological inorganic chemistry is a growing field of science that embraces the principles of biology and inorganic chemistry and impacts other fields ranging from medicine to the environment. JBIC (Journal of Biological Inorganic Chemistry) seeks to promote this field internationally. The Journal is primarily concerned with advances in understanding the role of metal ions within a biological matrix—be it a protein, DNA/RNA, or a cell, as well as appropriate model studies. Manuscripts describing high-quality original research on the above topics in English are invited for submission to this Journal. The Journal publishes original articles, minireviews, and commentaries on debated issues.