Synthesis And Performance Of A Novel Functional Material On Phosphorylated Protein Enrichment

Protein plays a vital role in the life events and is the direct manifestation of biological phenomena. Mechanisms in the physiological or pathological conditions need to be studied at the protein level. Meanwhile, the forms and the activities of the protein, for instance protein post-translational modification,interaction between proteins and protein conformation, depend on the study of the protein. Many proteins will have different modifications on the different groups after translated,such as phosphorylation. The selective and reversible phosphorylation of proteins is a key regulatory mechanism for biological processes, illustrated by the fact that30-50%of proteins might be phosphorylated at any time.In recent years, mass spectrometry (MS) has become an increasingly viable alternative to more traditional methods of phosphorylation analysis. However, a diversity of phosphorylated protein forms, and the transient nature of phosphorylation states, the amount of phosphorylated species in biological samples and protein digests is low because of a low phosphorylation stoichiometry. Therefore, it is necessary to develop efficient separation and enrichment methods for the detection of phosphorylated protein.In this study, we synthesised SiO2according to Stober Method and modified double-bonded hydrocarbon on the surface of SiO2as carrier. Meanwhile, we also synthesised,1-(4-Vinylphenyl)-3-(3,5-bis(trifluromethyl)phenyl)-urea,one kind of anion receptor as functional monomer and a kind of polymer material with cross-linker MBA and initiator ABDV on the surface of SiO2.lt named SiO2@M2which could absorption phosphorylation proteins specificallly. SiO2@M2was characterized by FT-IR and TEM respectively. The results indicated that SiO2@M2had a mean size of210-220nm in diameter, the thickness of polymer material with fuctional monomer was about10nm. Using β-Casein, the recognition properties of the obtained materials were evaluated. The adsorption results showed that SiO2@M2not only had a high absorption capacity of β-Casein(Q=92mg/g), but also had high absorption capacity of other two phosphorylated proteins, a-Casein (Q=40.2mg/g) and OVA(Q=36.3mg/g),which are higher than the absorption capacity of non-phosphorylated proteins,BSA(Q=12.8mg/g、Cyt c(6.28mg/g)、 BHb(3.8mg/g)、Lyz(8.08mg/g) respectively.We could also prove that SiO2@M2can selectively absorp and enrich phosphorylated proteins from proteins mixture systerm with SDS-PAGE protein gel electrophoresis.The SiO2@M2was also used to selectively recognize and enrich phosphorylated proteins from diluted skim milk. The results indicated that SiO2@M2displayed high selectivity.