State Key Laboratory of High-efficiency Utilization of Coal and Green Chemical Engineering, National Demonstration Center for Experimental Chemistry Education, School of Chemistry and Chemical Engineering, Ningxia University, Yinchuan 750021, China.
State Key Laboratory of High-efficiency Utilization of Coal and Green Chemical Engineering, National Demonstration Center for Experimental Chemistry Education, School of Chemistry and Chemical Engineering, Ningxia University, Yinchuan 750021, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 May 30;1199:123235. doi: 10.1016/j.jchromb.2022.123235. Epub 2022 Apr 5.
In this paper, we developed a facile route for the preparation of a novel bimetal oxide affinity chromatography (MOAC) material. The TiO/ZrO@MoS was constructed by the electrostatic interaction between titanium oxide/zirconia (w:w, 10:1) and molybdenum disulfide nanosheet. The nanocomposite has the large specific surface area (186.30 m⋅g) and pore volume (0.37 cm⋅g). Compared with single-metal probes, the combination of bimetallic oxides probe (TiO/ZrO) and hydrophilicity MoS support offered multitudinous affinity sites for phosphopeptides capturing from tryptic digests of protein samples under 50% acetonitrile-1% trifluoroacetate conditions. Singnificant feasibility of the TiO/ZrO@MoS nanomaterial for the enrichment of phosphopeptides under optimal conditions was proved via the bovine serum albumin (BSA) and the mixtures of β-casein. The phosphopeptide expression was identified using ultra-performance liquid chromatography (uHPLC) separation and-linear ion trap mass spectrometry (MS). With these affinity characters of TiO/ZrO@MoS, it exhibited higher binding capacity (25 mg⋅g), better selectivity for phosphopeptides from β-casein/BSA (1:2000) tryptic digests, high sensitivity (1 fmol⋅µL) towards phosphopeptides from β-casein tryptic digests, and great reusability of 8 cycles test for capturing phosphopeptides. In addition, the TiO/ZrO@MoS with high sensitivity and selectivity was successfully applied to enriching phosphopeptides from nonfat milk and human serum samples. More importantly, the TiO/ZrO@MoS was further successfully applied to multi-phosphopeptides enrichment, 1779 serine, threonine and tyrosine phosphosites can be identified in A549 cell protein tryptic digest. Compared with commercial TiO from enrichening 416 phosphopeptide from A549 cell lysates, the successful locating of 44 phosphosites were overlapped.
本文开发了一种简便的制备新型双金属氧化物亲和色谱（MOAC）材料的方法。TiO/ZrO@MoS 通过氧化钛/氧化锆（w:w，10:1）与二硫化钼纳米片之间的静电相互作用构建。该纳米复合材料具有较大的比表面积（186.30 m·g-1）和孔体积（0.37 cm·g-1）。与单金属探针相比，双金属氧化物探针（TiO/ZrO）和亲水性 MoS 载体的结合为在 50%乙腈-1%三氟乙酸条件下从蛋白质样品的胰蛋白酶消化物中捕获磷酸肽提供了众多亲和位点。通过牛血清白蛋白（BSA）和β-酪蛋白混合物，证明了 TiO/ZrO@MoS 纳米材料在最佳条件下用于富集磷酸肽的显著可行性。使用超高效液相色谱（uHPLC）分离和线性离子阱质谱（MS）鉴定了磷酸肽的表达。基于 TiO/ZrO@MoS 的这些亲和特性，它表现出较高的结合能力（25 mg·g-1），对β-酪蛋白/BSA（1:2000）胰蛋白酶消化物中的磷酸肽具有更好的选择性，对β-酪蛋白胰蛋白酶消化物中的磷酸肽具有较高的灵敏度（1 fmol·μL-1），并能重复使用 8 次循环测试来捕获磷酸肽。此外，TiO/ZrO@MoS 具有高灵敏度和选择性，成功应用于从脱脂牛奶和人血清样品中富集磷酸肽。更重要的是，TiO/ZrO@MoS 进一步成功地应用于多磷酸肽的富集，在 A549 细胞蛋白胰蛋白酶消化物中可以鉴定到 1779 个丝氨酸、苏氨酸和酪氨酸磷酸化位点。与从 A549 细胞裂解物中富集 416 个磷酸肽的商业 TiO 相比，成功定位了 44 个磷酸化位点。
