Proteomic analysis of posttranslational modification on topoisomerase 1 and centromere protein in patients with Scleroderma.

2014 Fiscal Year Final Research Report

• Project/Area Number: 25860820
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Collagenous pathology/Allergology
• Research Institution: St. Marianna University School of Medicine
• Principal Investigator: IIZUKA Nobuko 聖マリアンナ医科大学, 医学部, 講師 (00348533)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 強皮症 / プロテオミクス / 翻訳後修飾 / 自己抗原タンパク質

Outline of Final Research Achievements

To investigate the posttransratinal modification on Topoisomerase Ⅰ (topo1) and centromere B protein (CENTB) which are autoantigenic proteins of Scleroderma.
We separated proteins extracted from peripheral blood mononuclear cells (PBMCs) from 3 patients and healthy donors to 2-dimentional electrophoresis and performed western blotting (2DE-WB) using anti-topo1 antibodies and anti-CENTB antibodies, respectively. We analyzed the differently reactivities of protein spots on 2DE-WB between the screroderma (Ssc) and the healthy contorol (HC) groups.
In 2DE-WB of topo1 analysis, we detected 12 protein spots, and 5 out of 12 spots have more than 2-times stronger or lower reactivities in Ssc than in HC groups. Also in 2DE-WB of CENTB analysis, we detected 8 protein spots and 4 out of 8 spots have more than 2-times stronger reactivities in Ssc than HC groups. We tried to identify these proteins, which have differential reactivities in 2DE-WB between Ssc and HC groups, by mass spectrometry.

Free Research Field

膠原病