Hydrophilic interaction liquid chromatography: An efficient tool for assessing thorium interaction with phosphorylated biomimetic peptides

• Published in: Journal of Chromatography A Vol. 1735; p. 465341
• Main Authors: Abou-Zeid, Lana, Pell, Albert, Amaral Saraiva, Marina, Delangle, Pascale, Bresson, Carole
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25.10.2024; Elsevier
• Subjects: Affinity; Chemical Sciences; Competition; ESI-MS; HILIC; ICP-MS; Phosphorylated peptides; Thorium; Competition; Phosphorylated peptides; Affinity; ESI-MS; ICP-MS; HILIC; Thorium; Thorium Phosphorylated peptides Affinity Competition HILIC ESI
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2024.465341

•HILIC has proven to be efficient to achieve separation of actinide-complexes.•First HILIC separations of thorium-phosphorylated peptide complexes are reported.•The dimeric structure of the new complexes was preserved through HILIC separation.•Th quantitative distribution in the complexes was determined by HILIC-ESI-MS/ICP-MS.•Th4+ interacts more strongly than UO22+ with the tetraphosphorylated peptide pS1368. In the field of nuclear toxicology, the knowledge of the interaction of actinides (An) with biomolecules is of prime concern in order to elucidate their toxicity mechanism and to further develop selective decorporating agents. In this work, we demonstrated the great potential of hydrophilic interaction liquid chromatography (HILIC) to separate polar thorium (Th) biomimetic peptide complexes, as a key starting point to tackle these challenges. Th4+ was used as plutonium (Pu4+) analogue and pS16 and pS1368 as synthetic di- and tetra-phosphorylated peptides capable of mimicking the interaction sites of these An in osteopontin (OPN), a hyperphosphorylated protein. The objective was to determine the relative affinity of pS16 and pS1368 towards Th4+, and to evaluate the pS1368 selectivity when Th4+ was in competition complexation reaction with UO22+ at physiological pH. To meet these aims, HILIC was simultaneously coupled to electrospray ionization mass spectrometry (ESI-MS) and inductively coupled plasma mass spectrometry (ICP-MS), which allowed to identify online the molecular structure of the separated complexes and quantify them, in a single step. Dedicated HILIC conditions were firstly set up to separate the new dimeric Th2(peptide)2 complexes with good separation resolution (peptide = pS16 or pS1368). By adding pS16 and pS1368 in different proportions relatively to Th4+, we found that lower or equal proportions of pS16 with respect to pS1368 were not sufficient to displace pS1368 from Th2pS13682 and pS16 proportion higher than pS1368 led to the formation of a predominant ternary complex Th2(pS16)(pS1368), demonstrating preferential Th4+ binding to the tetra-phosphorylated peptide. Finally, online identification and quantification of the formed complexes when Th4+ and UO22+ were mixed in equimolar ratio relatively to pS1368 showed that in spite of pS1368 has been specifically designed to coordinate UO22+, pS1368 is also Th4+-selective and exhibits stronger affinity for this latter than for UO22+. Hence, the results gathered through this approach highlight the impact of Th4+ coordination chemistry on its interaction with pS1368 and more widely to its affinity for biomolecules.