Absolute Quantification of Transmembrane Protein Shedding and Its Application to the Monitoring of Chronic Inflammation

2013 Fiscal Year Final Research Report

• Project/Area Number: 24659280
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Laboratory medicine
• Research Institution: Ehime University
• Principal Investigator: HIGASHIYAMA Shigeki 愛媛大学, プロテオサイエンスセンター, 教授 (60202272)
• Co-Investigator(Renkei-kenkyūsha): MIYAZAKI Tatsuhiko 岐阜大学, 医学部附属病院病理部, 臨床教授 (80239384) ; TAKEMORI Nobuaki 愛媛大学, プロテオサイエンスセンター, 講師 (40533047)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: 臨床検査システム / shedding / タンパク質絶対定量 / プロテオーム解析

Research Abstract

Ectodomain shedding of transmembrane proteins is induced by inflammatory factors such as chemokines and cytokines. In order to monitor the progress of chronic inflammation, we attempted to establish a monitoring system of the ectodomain shedding using the combination of an originally synthesized internal standard protein mixture and MRM by mass spectrometry. Then, we successfully accomplished the absolute measurement of transmembrane protein contents in mouse tissues at f-mol levels.

Research Products

• [Journal Article] Cisplatin resistance by induction of aldo-keto reductase family 1 member C2 in human bladder cancer cells (2014)
  • Author(s): Shirato A, Kikugawa T, Miura N, Tanji N, Takemori N, Higashiyama S, Yokoyama M.
  • Journal Title: Oncol Lett Volume: 7 Pages: 674-678
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/24527071
  • Peer Reviewed
• [Journal Article] Annexin A2 regulates a disintegrin and metalloproteinase 17-mediated ectodomain shedding of pro-tumor necrosis factor-αin monocytes and colon epithelial cells (2013)
  • Author(s): T sukamoto H, Tanida S, Ozeki K, Ebi M, Mizoshita T, Shimura T, Mori Y, Kataoka H, Kamiya T, Fukuda S, Higashiyama S, Joh T.
  • Journal Title: Inflamm Bowel Dis Volume: 19(7) Pages: 1365-73
  • DOI: 10.1097/MIB.0b013e318281f43a
  • Peer Reviewed
• [Journal Article] Spatiotemporal visualization of proHB-EGF ectodomain shedding in living cells (2013)
  • Author(s): Inoue H, Sakaue T, Ozawa T, Higashiyama S.
  • Journal Title: J Biochem Volume: 154 Pages: 67-76
  • DOI: 10.1093/jb/mvt030
  • Peer Reviewed
• [Journal Article] Notch increases the shedding of HB-EGF by ADAM12 to potentiate invadopodia formation in hypoxia (2013)
  • Author(s): Díaz B, Yuen A, Iizuka S, Higashiyama S, Courtneidge SA.
  • Journal Title: J Cell Biol Volume: 201 Pages: 279-292
  • DOI: 10.1083/jcb.201209151
  • Peer Reviewed
• [Journal Article] Cisplatin influences acquisition of resistance to molecular-targeted agents through epithelial-mesenchymal transition-like changes (2013)
  • Author(s): Kurokawa M, Ise N, Omi K, Goishi K, Higashiyama S.
  • Journal Title: Cancer Sci Volume: 104 Pages: 904-911
  • DOI: 10.1111/cas.12171
  • Peer Reviewed
• [Journal Article] Telmisartan inhibits cell proliferation by blocking nuclear translocation of ProHB-EGF C-terminal fragment in colon cancer cells (2013)
  • Author(s): Ozeki K, Tanida S, Morimoto C, Inoue Y, Mizoshita T, Tsukamoto H, Shimura T, Kataoka H, Kamiya T, Nishiwaki E, Ishiguro H, Higashiyama S, Joh T.
  • Journal Title: PLoS One Volume: 8 Pages: e56770
  • DOI: 10.1371/journal.pone.0056770
  • Peer Reviewed
• [Journal Article] Human antigen R-mediated mRNA stabilization is required for ultraviolet B-induced autoinduction of amphiregulin in keratinocytes (2013)
  • Author(s): Nakayama H, Fukuda S, Matsushita N, Nishida-Fukuda H, Inoue H, Shirakata Y, Hashimoto K, Higashiyama S.
  • Journal Title: J Biol Chem Volume: 288 Pages: 10338-10348
  • DOI: 10.1074/jbc.M112.417527
  • Peer Reviewed
• [Journal Article] TGFαshedding assay : an accurate and versatile method for detecting GPCR activation (2012)
  • Author(s): Inoue A, Ishiguro J, Kitamura H, Arima N, Okutani M, Shuto A, Higashiyama S, Ohwada T, Arai H, Makide K, Aoki J.
  • Journal Title: Nat Methods Volume: 9 Pages: 1021-1029
  • DOI: 10.1038/nmeth.2172
  • Peer Reviewed
• [Journal Article] Cell surface annexins regulate ADAM-mediated ectodomain shedding of proamphiregulin (2012)
  • Author(s): Nakayama H, Fukuda S, Inoue H, Nishida-Fukuda H, Shirakata Y, Hashimoto K, Higashiyama S.
  • Journal Title: Mol Biol Cell Volume: 23 Pages: 1964-1975
  • DOI: 10.1091/mbc.E11-08-0683
  • Peer Reviewed
• [Journal Article] A disintegrin and metalloenzyme (ADAM) 17 activation is regulated byα5β1 integrin in kidney mesangial cells (2012)
  • Author(s): Gooz P, Dang Y, Higashiyama S, Twal WO, Haycraft CJ, Gooz M.
  • Journal Title: PLoS One Volume: 7 Pages: e33350
  • DOI: 10.1371/journal.pone.0033350
  • Peer Reviewed
• [Presentation] Absolute Quantitative Analysis of Urinary Proteome Alterations Using Wheat Germ Cell Free Protein Synthesis and MRM Mass Spectrometry (2013)
  • Author(s): Takemori N, Takemori A, Higashiyama S.
  • Organizer: HUPO 11th Annual World Congress
  • Place of Presentation: Boston, MA, USA
  • Year and Date: 20130909-13
• [Presentation] Ectodomain shedding制御とEGFR/Ligandシグナル (2013)
  • Author(s): 東山繁樹、福田信二
  • Organizer: 第18回日本病態プロテアーゼ学会
  • Place of Presentation: 大阪
  • Year and Date: 20130816-17
• [Presentation] コムギ胚芽無細胞タンパク質合成系を用いた絶対定量プロテオーム解析技術の開発 (2013)
  • Author(s): 武森信暁、武森文子、森下了、青島理人、澤崎達也、東山繁樹
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: 愛知
  • Year and Date: 20130619-21
• [Presentation] Ectodomain shedding in EGFR/ligand signaling; regulation, functional implication and imaging. Matrix Metalloproteinases (2013)
  • Author(s): Higashiyama S.
  • Organizer: Gordon Research Conference
  • Place of Presentation: Lucca, Italy
  • Year and Date: 20130519-24
• [Remarks]
  • URL: http://www.m.ehime-u.ac.jp/data/course/list_detail.php?id=201100000053