An integrated method for the detection of basic proteins in serum-derived proteomes. Issue 8 (August 2016)

• Record Type: Journal Article
• Authors: Yin, Shang-Jun; Yang, Hee Seung; Cho, Ick-Hyun; Wang, Qian; Park, Yong-Doo; Yang, Jun-Mo
• Is Part Of: Process biochemistry. Volume 51:Issue 8(2016:Aug.)
• Page Start: 973
• Page End: 980
• Publication Date: 2016-08
• Subjects: AD atopic dermatitis -- ADe extrinsic type of AD -- 2D-PAGE two-dimensional polyacrylamide gel electrophoresis -- MALDI-TOF matrix-assisted laser desorption/ionization-time of flight -- IPG immobilized pH gradient -- MARC multiple affinity removal column -- IEF isoelectric focusing; Serum proteomics -- Basic proteins -- Serum fractionation -- Integration -- Atopic dermatitis; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2016.04.023
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: The difficulty of serum proteomics application for the basic regions of proteins. Integrating approach by using Micro-Rotofor IEF fractionation and AU-PAGE. Successful detections for C3 and C4A complement fragments proteins. Basic protein detection in atopic dermatitis patients-derived sera samples. Abstract: Because serum is an optimal source to identify proteins derived from diseased-tissue compartments, serum proteomics has been applied for the discovery and analysis of biomarkers related to human disease. The general 2D-PAGE method is suitable for acidic and neutral proteome separation while highly basic proteins remain unresolved. In this study, we optimized basic proteome fractionation by integrating several methods such as, Micro-Rotofor isoelectric focusing (IEF) fractionation, acetic acid-urea polyacrylamide gel electrophoresis (A-U PAGE) separation, and mass spectrometry analysis for identifying basic proteins in patient sera. The sera samples were obtained from patients with atopic dermatitis (AD) to establish the model method introduced in the present study. We successfully identified the specific presence of a C3 complement component fragment protein and C4A fragment protein in AD patient-derived sera. Highly basic proteins in serum are very difficult to detect due to their separation properties. Thus, the integrated proteomic method approaches described here could be applicable for the detection of basic proteins associated with other diseases.