Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, Pakistan.
Anal Chem. 2012 Dec 4;84(23):10180-5. doi: 10.1021/ac3023197. Epub 2012 Nov 16.
Relying on the successful journey of metal oxides in phosphoproteomics, lanthanum oxide is employed for the engineering of an affinity material for phosphopeptide enrichment. The lanthanum oxide is chemically modified on the surface of silica and characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and Fourier transform infrared spectroscopy (FTIR). The obtained silica-lanthanum oxide composite is applied for the selective enrichment of phosphopeptides from tryptic digest of standard protein (α-casein, β-casein, and commercially available casein mixtures from bovine milk). The enriched entities are analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The mass spectroscopy (MS) results show that the silica-lanthanum oxide composite exhibits enhanced capability for phosphopeptide enrichment with sensitivity assessed to be 50 fmol. Sequence coverage of casein is interpreted showing successful recovery. As a real sample, a protein digest of nonfat milk is applied. Also, the ability of lanthanum in different formats is checked in the selective phosphopeptides enrichment. The composite holds promising future in economic ground as it also possesses the regenerative ability for repetitive use.
基于金属氧化物在磷酸化蛋白质组学中取得的成功经验,氧化镧被用于设计用于磷酸肽富集的亲和材料。氧化镧在二氧化硅表面进行化学修饰,并通过扫描电子显微镜(SEM)、能谱(EDX)和傅里叶变换红外光谱(FTIR)进行表征。所得的二氧化硅-氧化镧复合材料用于从标准蛋白(α-酪蛋白、β-酪蛋白和市售牛乳酪蛋白混合物)的胰蛋白酶消化物中选择性富集磷酸肽。通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)对富集物进行分析。质谱(MS)结果表明,二氧化硅-氧化镧复合材料在磷酸肽富集方面表现出增强的能力,其灵敏度评估为 50 fmol。对酪蛋白的序列覆盖进行解释,表明成功回收。作为实际样品,应用了非脂乳蛋白的蛋白质消化物。此外,还检查了不同形式的镧在选择性磷酸肽富集中的能力。由于该复合材料还具有重复使用的再生能力,因此在经济方面具有广阔的应用前景。