Document Name: Quantifying Protein-Specific N-Glycome Profiles by Focused Protein and Immunoprecipitation Glycomics (focus and glycomics Quantitative immunoprecipitation N- glycosylation pattern)
Journal Name: Journal of Proteome Research
Published: 2019 Aug 2
DOI: 10.1021/acs.jproteome.9b00232
IF: 4.173
unit:
1, Japan and Hepatology Department of the Graduate School of Medicine, Hokkaido University, Sapporo gastrointestinal
2, Sapporo City, Japan Ishinomaki pharmaceutical research and innovation center
3, Hokkaido University in Sapporo, Japan Advanced Institute of Life Science and Medicine Laboratory functional sugar group
Experimental Materials: The five non-alcoholic fatty liver (nonalcoholic fatty liver; NAFL) 6 NON - patients with alcoholic hepatitis (nonalcoholic steatohepatitis; NASH) serum
Technology: Focus glycoproteins assay (focused protein glycomics; FPG), glycomics immunoprecipitation (immunoprecipitation glycomics; IPG) combined MALDI-TOF MS technique.
Share People: Jennifer Pan Fire
I. Overview:
In the previous study, a method of using FPG combined MALDI-TOF MS on hepatitis model mouse serum N- glycosylation quantitative analysis, found that some N- glycosylation in development of significant changes in hepatitis; although the overall protein FPG method Keyihuode N- glycosylation pattern specific, but the search for candidate markers in the more samples, FPG method of operating a cumbersome affinity separation, in-gel digested protein extracts, the time-consuming. FPG method for limitations, the authors developed a new method for IPG. IPG method is the use of affinity beads with protein immunoaffinity precipitation reaction, combined techniques MALDI-TOF MS analysis of sugars.
Comparative herein FPG mainly two methods IPG and human serum samples nonalcoholic fatty liver disease α-1 antitrypsin (alpha-1 antitrypsin, ATT) N- quantitative sugar level and ceruloplasmin (ceruloplasmin, CP), with the result IPG these methods and found that N- glycosylation quantitative FPG two proteins having good correlation; and, IPG and FPG method has good correlation is obtained on the ATT and CP sugar. FPG described IPG and method can achieve quantitative analysis of the sugars N-. IPG herein by FPG and methods were found ATT-A3F increased most notably in the non-alcoholic hepatitis, for this estimation may ATT-A3F nonalcoholic hepatitis candidate biomarkers.
Second, the background:
Current clinical diagnosis is dependent on NALD biopsy, biopsy is not only invasive large, complicated operation, and there is also sampling bias. To do this, find an effective noninvasive biomarker has a more important significance.
Glycosylation is one of the most common post-translational modification of proteins, many studies have shown that changes in serum protein N- glycans may be a diagnostic marker for liver-related diseases. Therefore, the analysis of serum protein specific N- Sugar is a powerful strategy to find new biomarkers of liver disease.
Third, the experimental design:
In this paper, FPG and IPG serum five nonalcoholic fatty liver disease and six non-alcoholic hepatitis patients were N- sugar analysis, in order to find new non-alcoholic hepatitis biomarkers through. First, using the method of FPG protein selected 7 N- glycosylation analysis, screened two most obvious change ATT and the results of CP protein; IPG then followed using these two methods of screening proteins N- glycosylation analysis.
Fourth, the research results:
Human serum albumin 4.1 glycomics analysis scheme N-
4.2 FPG N- glycosylation for histological analysis 7 group selected proteins
4.3 FPG 7 Analysis of selected serum glycoprotein information list
4.4 FPG N- glycosylation pattern analysis result of the measurement of selected serum proteins 7
4.5 IPG FPG and has good correlation to quantification ATT-A3F
4.6 IPG Determination of α-1 antitrypsin N- glycosylation pattern
4.7 FPG is obtained and the amount of IPG ATT-A3F sugar chain and CP-A2 correlation
V. Conclusion:
FPG and methods can IPG N- glycosylation by analysis to find biomarkers of disease, FPG level suitable for the N- glycosylation comprehensive analysis of protein gel isolated, when applied to IPG samples N- large number of sugar analysis. We find ATT-A3F may nonalcoholic hepatitis biomarkers.