Kupcik Rudolf, Macak Jan M, Rehulkova Helena, Sopha Hanna, Fabrik Ivo, Anitha V C, Klimentova Jana, Murasova Pavla, Bilkova Zuzana, Rehulka Pavel
Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech Republic.
Center of Materials and Nanotechnologies, Faculty of Chemical Technology, University of Pardubice, Nam. Cs. Legii 565, 530 02 Pardubice, Czech Republic.
ACS Omega. 2019 Jul 15;4(7):12156-12166. doi: 10.1021/acsomega.9b00571. eCollection 2019 Jul 31.
This work reports highly selective phosphopeptide enrichment using amorphous TiO nanotubes (TiONTs) and the same material decorated with superparamagnetic FeO nanoparticles (TiONTs@FeONPs). TiONTs and TiONTs@FeONPs materials were applied for phosphopeptide enrichment both from a simple peptide mixture (tryptic digest of bovine serum albumin and α-casein) and from a complex peptide mixture (tryptic digest of Jurkat T cell lysate). The obtained enrichment efficiency and selectivity for phosphopeptides of TiONTs and TiONTs@FeONPs were increased to 28.7 and 25.3%, respectively, as compared to those of the well-established TiO microspheres. The enrichment protocol was extended for a second elution step facilitating the identification of additional phosphopeptides. It further turned out that both types of amorphous TiO nanotubes provide qualitatively new physicochemical features that are clearly advantageous for highly selective phosphopeptide enrichment. This has been confirmed experimentally resulting in substantial reduction of non-phosphorylated peptides in the enriched samples. In addition, TiONTs@FeONPs combine high selectivity and ease of handling due to the superparamagnetic character of the material. The presented materials and performances are further promising for applications toward a whole range of other types of biomolecules to be treated in a similar fashion.
本研究报道了使用非晶态TiO纳米管(TiONTs)以及用超顺磁性FeO纳米颗粒修饰的相同材料(TiONTs@FeONPs)进行高选择性磷酸肽富集。TiONTs和TiONTs@FeONPs材料被用于从简单肽混合物(牛血清白蛋白和α-酪蛋白的胰蛋白酶消化物)以及复杂肽混合物(Jurkat T细胞裂解物的胰蛋白酶消化物)中富集磷酸肽。与成熟的TiO微球相比,TiONTs和TiONTs@FeONPs对磷酸肽的富集效率和选择性分别提高到了28.7%和25.3%。富集方案扩展到了第二个洗脱步骤,有助于鉴定更多的磷酸肽。进一步发现,这两种类型的非晶态TiO纳米管都提供了全新的物理化学特性,这对于高选择性磷酸肽富集显然是有利的。这已通过实验得到证实,富集样品中未磷酸化肽的数量大幅减少。此外,由于材料的超顺磁性,TiONTs@FeONPs兼具高选择性和易于处理的特点。所展示的材料和性能对于以类似方式处理的一系列其他类型生物分子的应用具有进一步的前景。