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

• 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
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• 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 / プロテオーム解析 / トポイソメラーゼI / セントロメア

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.

Research Products

• [Journal Article] Effect of sulfasalazine and tofacitinib on the protein profile of articular chondrocytes. (2014)
  • Author(s): Endo W, Arito M, Sato T, Kurokawa MS, Omoteyama K, Iizuka N,Okamoto K, Suematsu N, Nakamura H, Beppu M, Kato T.
  • Journal Title: Modern Rheumatology Volume: 24 Pages: 844-850
  • Peer Reviewed
• [Journal Article] Proteomic analysis of whole glomeruli in patients with IgA nephropathy using microsieving. (2014)
  • Author(s): Kojima S, Koitabashi K, Iizuka N, Okamoto K, Arito M, Sato T, Kurokawa MS, Suematsu N, Shibagaki Y, Yasuda T, Kimura K, Kato T
  • Journal Title: American journal of nephrology. Volume: 39 Issue: 1 Pages: 36-45
  • DOI: 10.1159/000357788
  • Peer Reviewed
• [Journal Article] Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis (2013)
  • Author(s): Kurokawa MS, et al
  • Journal Title: J Proteomics Volume: 91C Pages: 259-69
  • DOI: 10.1016/j.jprot.2013.07.021
  • NAID: 130005454504
  • Peer Reviewed
• [Journal Article] Protein profiles of peripheral blood mononuclear cells as a candidate biomarker for Behcet’s disease. (2013)
  • Author(s): Yoshioka T, Kurokawa MS, Sato T, Nagai K, Iizuka N, Arito M, Takakuwa Y, Nakano H, Ooka S, Suematsu N, Okamoto K, Yudoh K, Nakamura H, Suzuki N, Ozaki S, Kato T.
  • Journal Title: Clinical and Experimental Rheumatology, Volume: Epub
  • Peer Reviewed
• [Journal Article] Roles of serum fibrinogen α chain-derived peptides in Alzheimer's disease. (2013)
  • Author(s): Noguchi M, Sato T, Nagai K, Utagawa I, Suzuki I, Arito M, Iizuka N, Suematsu N, Okamoto K, Kato T, Yamaguchi N, Kurokawa MS.
  • Journal Title: International Journal of Geriatric Psychiatry Volume: in press Issue: 8 Pages: 808-818
  • DOI: 10.1002/gps.4047
  • NAID: 130005454787
  • Peer Reviewed
• [Journal Article] Effect of sulfasalazine and tofacitinib on the protein profile of articular chondrocytes. (2013)
  • Author(s): Endo W, Arito M, Sato T, Kurokawa MS, Omoteyama K, Iizuka N,Okamoto K, Suematsu N, Nakamura H, Beppu M, Kato T.
  • Journal Title: Modern Rheum. Volume: Epub
  • Peer Reviewed
• [Presentation] Protein profiles of peripheral blood mononuclear cells as a biomarker for Behcet’s disease (2013)
  • Author(s): Yoshioka T, Kurokawa MS, Sato T, Nagai K, Iizuka N, Arito M, Takakuwa Y, Nakano H, Ooka H, Suematsu N, Okamoto K, Nakamura H, Suzuki N, Ozaki S, Kato T.
  • Organizer: HUPO2013
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
• [Presentation] Comparative croteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis. (2013)
  • Author(s): Uchida T, Nagai K, Sato T, Iizuka N, Arito M, Takakuwa Y, Nakano H, Ooka S, Kurokawa MS, Suematsu N, Okamoto K, Ozaki S, Kato T.
  • Organizer: HUPO2013
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
• [Presentation] Proteomic analysis of effects of antirheumatic drugs on exosomes derived from synovial sarcoma cells (2013)
  • Author(s): Tsuno H, Suematsu N, Sato T, Arito M, Iizuka N, Omoteyama K, Kurokawa MS, Okamoto K, Kato T.
  • Organizer: HUPO2013
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
• [Presentation] 2D-DIGE analysis of cell surface proteins reveals characteristic proteome profiles for effects of sulfasalazine in human synovial sarcoma cell line (2013)
  • Author(s): Omoteyama K, Arito M, Sato T, Iizuka N, Kurokawa MS, Okamoto K, Suematsu N, Kato T.
  • Organizer: HUPO2013
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
• [Presentation] Proteomic analysis of whole glomeruli in patients with IgA nephropathy using micro-sieving (2013)
  • Author(s): Kojima S, Koitabashi K, Iizuka N, Okamoto K, Arito M, Sato T, Kurokawa MS, Suematsu N, Yasuda T, Kimura K, Kato T.
  • Organizer: HUPO2013
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
• [Book] 最新関節リウマチ学 血清マトリックスメタプロテイナーゼ３ (2014)
  • Author(s): 飯塚進子
  • Total Pages: 5
  • Publisher: 日本臨床社