Purification And Identification Of Mannose-Binding Protein From Sera Of Health Human And HCC Patients

Background:Glycan binding proteins (GBPs) play a significant role in decoding the information content of glycans by recognizing and specifically binding to glycosylated protein and lipid ligands. As vital constituents of all living systems, GBPs are involved in recog-nition, adherence, motility, complement activation, and signaling processes. When the tumor occurs, the glycosylation of protein and lipid molecules change anomalies occurred caused glycan chains and number of structure changes, correspondingly the interaction of these glycan chains and GBPs also occur exceptionally change.In most cancer occurrence and development process the anomalies related GBPs type and quantity are also not know,the enormous complexity of GBP-ligand interactions and the absence of well defined glycans libraries and efficient analyticalscreening methods have limited analysis of their specificity and elucidation of their biological roles.In recent years, the existing problems in the liver tissue has been purified, through pregnancy-associated chain structure and study its glycans with normal liver cells, compared to the same glycoprotein found N-glycan chain structure changes, but the mechanism of these glycan chains interactions with protein expression is how to change, were know less. Liver cancer is a serious threat to human health diseases, how to effectively carry out the liver in combination with GBPs interaction system research, especially for hepatocellular carcinoma is closely related with the development of the GBPs separation, purification, identification and functionalization study for this disease is early diagnosis and treatment and related drug development is very significant. Based on the above, this paper adopts magnetic particles functional separation and purification of new technology in proteomics research comparatively mature method in two-dimensional electrophoresis technology and mass spectrometry technique to patients with hepatocellular carcinoma, coupled with normal serum of serum MBP separation, purification and appraisal analysis.Purpose:By using glycan magnetic particles complex purification out normal serum and MBP of liver cancer patients, then use 2D electrophoresis analysis and mass spectrometry appraisal hepatocellular carcinoma patients analysis, find out the differences between the serum and normal serum protein, the MBP specificity of liver cancer were analyzed. In order to find the liver cancer diagnosis potential molecular markers, for early diagnosis of HCC and resistance to liver cancer drug target molecules provide.Method:Use glycan magnetic particles complex purification out normal serum and liver cancer patients in the serum of mannose binding proteins (MBP), then a 2D electrophoresis analysis, MS appraisal (LC-MS/MS), searchable database of liver cancer patients, and normal mannose binding protein analysis, finally compared to liver cancer patients serum peculiar MBP for functionalization analysis and prediction.Result:we identified there having protein in serum healthy 75, protein in liver patient serum 79, health and liver cancer patients has 59 same protein, healthy human serum peculiar protein has 16, and liver cancer serum protein peculiar with 20, a total of 36difference protein. All proteins obtained analyzing data. The protein findings identify to 23 in http://www.uniprot.org database is no GO annotation,2 new protein in the database is not found.we founded six proteins, function is closely related with the occurrence of cancer, including two proteins can be considered as a potential of HCC diagnosed mannose combination of molecular markers. Quantity of proteins identified in this study can be estimated by Exponentially Modified Protein Abundance Index (emPAI). SAA1 serum amyloid A2 isoform a, Apolipoprotein E, Isoform 1 of Fibrinogen alpha chain, Isoform Gamma-B of Fibrinogen gamma chain, Isoform 1 of Fibronectin, Alpha-1B-glycoprotein, Isoform 2 of Inter-alpha-trypsin inhibitor heavy chain H4, Vitronectin, IGL@ protein, Putative uncharacterized proteinz were more than 1.5 fold up-regulated while Isoform 1 of Optineurin, Complement C1s subcomponent, Serum amyloid A-4 protein, Apolipoprotein A-â…¡, Apolipoprotein C-II, Transthyretin, CD5 antigen-like, Apolipoprotein M, Isoform LMW of Kininogen-1, Serum paraoxonase/arylesterase 1, cDNA FLJ54471, vitamin D-binding protein precursor were less than 0.66 fold down-regulated.Conclusion:This experiment using glycan magnetic particle composites purification out mannose binding protein,two-dimensional electrophoresis, mass spectrometry this set of experiment scheme hepatocellular carcinoma patients serum antifonding MBP peculiar to the MBP, conducted effectively appraisal and analysis, found six of protein in liver tumors, and there are two closely related to the MBP can serve as liver cancer diagnosis as potential markers and clinical diagnosis and treatment of HCC molecular targets. The results are proved the experiment method of image galleries of complex samples can be effective, high throughput, rapid separation, purification, appraisal and analysis. And using emPAI index analysis method for the identification of the MBP to quantitatively analyzed with protein of Health and liver cancer were identified differential expression of serum mannose binding protein...