Characterization and applications of enzymatic cross-linking reactions

• Project/Area Number: 20200072
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
• Allocation Type: Single-year Grants
• Research Field: Applied biochemistry; Applied molecular and cellular biology
• Research Institution: Nagoya University
• Principal Investigator: KIYOTAKA Hitomi 名古屋大学, 生命農学研究科, 准教授 (00202276)
• Co-Investigator(Renkei-kenkyūsha): KAMIYA Noriho 九州大学, 大学院・工学系研究科, 教授 (50302766) ; MTSUFUJI Senya 東京慈恵会医科大学, 医学部, 教授 (50192753)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥33,800,000 (Direct Cost: ¥26,000,000、Indirect Cost: ¥7,800,000); Fiscal Year 2010: ¥11,050,000 (Direct Cost: ¥8,500,000、Indirect Cost: ¥2,550,000); Fiscal Year 2009: ¥11,050,000 (Direct Cost: ¥8,500,000、Indirect Cost: ¥2,550,000); Fiscal Year 2008: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
• Keywords: 酵素 / 基質 / カルシウム / 蛋白質接着 / 基質配列 / トランスグルタミナーゼ / タンパク質架橋 / 翻訳後修飾 / 表皮

Research Abstract

Enzymatic cross-linking is essential reaction for several biological events: blood coagulation, skin formation. Transglutaminase, which is responsible for the reaction, is a family of enzyme consisting of 8 isozymes. In this project, I have carried out the identification of highly reactive substrate peptide which is useful tool for resolving of physiological significance. Furthermore, application of the peptide has been attempted.

Research Products

• [Journal Article] Knocking-in the R142C mutation in transglutaminase 1 disrupts the stratum comeum barrier and postnatal survival of mice (2012)
  • Author(s): Nakagawa N, Yamamoto M., Hitomi K, (他 10名)Yamanishi K.
  • Journal Title: J. Dermatol. Sci. Volume: 65 Pages: 192-206
  • Peer Reviewed
• [Journal Article] Knocking-in the R142C mutation in transglutaminase 1 disrupts the stratum corneum barrier and postnatal survival of mice (2012)
  • Author(s): Nakazawa N, Yamamoto M., Hitomi K, Yamanishi K., 他10名
  • Journal Title: Journal of Dermatological Science Volume: 65 Issue: 3 Pages: 196-206
  • DOI: 10.1016/j.jdermsci.2011.12.011
  • Peer Reviewed
• [Journal Article] Hornerin is a component of the epidermal comified cell envelopes (2011)
  • Author(s): Henry J, Hitomi K., Simon M., (他8名)
  • Journal Title: FASEB J. Volume: 25(5) Pages: 1567-1576
  • Peer Reviewed
• [Journal Article] Screening of substrate peptide sequences for tissue-type transglutaminase (TGase 2) Using T7 Phage cDNA Library (2011)
  • Author(s): Sugimura, Y. Yamashita H. Hitomi K.
  • Journal Title: Cytotechnology Volume: 63(2) Pages: 111-118
  • Peer Reviewed
• [Journal Article] In situ detection of active transglutaminases for keratinocyte-type (TGase 1) and tissue-type (TGase 2) using fluorescence-labeled highly reactive substrate peptides (2011)
  • Author(s): Itoh M., Kawamoto T., Tatsukawa H., Kojima S., Yamanishi, K., Hitomi. K.
  • Journal Title: J. Histochem. Cytochem. Volume: 59 Pages: 180-187
  • Peer Reviewed
• [Journal Article] Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles (2011)
  • Author(s): Yamane, A., Fukui, M., Sugimura, Y., Itoh, M., Alea, M. P, Said, El Alaoui, Akiyama, M., Hitomi, K.
  • Journal Title: FEBS J. Volume: 277 Pages: 3564-3574
  • Peer Reviewed
• [Journal Article] In situ detection of active transglutaminases for keratinocyte-type (TGase 1) and tissue-type (TGase 2) using fluorescence-labeled highly reactive substrate peptides (2011)
  • Author(s): Itoh, H, Kawamoto T., Hitomi K, 他3名
  • Journal Title: Journal of Histochemistry and Cytochemistry Volume: 59 Pages: 180-187
  • Peer Reviewed
• [Journal Article] Screening of substrate peptide sequences for tissue-type transglutaminase (TGase 2) Using T7 Phage cDNA Library (2011)
  • Author(s): Sugimura. Y, Yamashita, K., Hitomi K.
  • Journal Title: Cytotechnology Volume: 59 Pages: 180-187
  • Peer Reviewed
• [Journal Article] Hornerin is a component of the epidermal comified cell envelopes (2011)
  • Author(s): Henry, J, Hitomi, K, Simon M, 他10名
  • Journal Title: FASEB Journal Volume: 25 Issue: 5 Pages: 1567-1576
  • DOI: 10.1096/fj.10-168658
  • Peer Reviewed
• [Journal Article] The cystatin M/E-cathepsin L balance is essential for tissue homeostasis in epidermis, hair follicles, and cornea (2010)
  • Author(s): Zeeuwen PM., van Vlijmen-Willems I., Rodijk-Olthuisl, D., Hitomi K., Schalkwijk J., (他5名)
  • Journal Title: FASEB J. Volume: 24 Pages: 3744-3755
  • Peer Reviewed
• [Journal Article] Transglutaminase 1 Preferred Substrate Peptide K5 Is an Efficient Tool in Diagnosis of Lamellar Ichthyosis (2010)
  • Author(s): Akiyama M., Hitomi K. and Shimizu, H., (他4名)
  • Journal Title: Am. J. Pathol. Volume: 129 Pages: 2306-2309
  • Peer Reviewed
• [Journal Article] Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles (2010)
  • Author(s): Yamane, A., Fukui M.Hitomi, et al.(他5名)
  • Journal Title: FEBS Journal Volume: 277 Pages: 3564-3574
  • Peer Reviewed
• [Journal Article] Transglutaminase 1 preferred substrate peptide K5 is an efficient tool in diagnosis of lamellar ichthyosis. (2010)
  • Author(s): Akiyama M, Sakai K, Teruki K, Yanagi T, Fukushima S, Ihn H, Hitomi K, Shimizu, H
  • Journal Title: American Journal Pathology 129 Pages: 306-2309
  • Peer Reviewed
• [Journal Article] A Specific Colorimetric assay for measuring of transglutaminase 1 and factor XIII activities (2009)
  • Author(s): Hitomi, K., Kitamura, M., Alea, M. P., Ismail, C., Thomas, V., Alaoui, S El.
  • Journal Title: Anal. Biochem. Volume: 394 Pages: 281-283
  • Peer Reviewed
• [Journal Article] A biochemical pathway that controls skin barrier formation: expression of its key components in inflammatory skin diseases and in reconstructed skin (2009)
  • Author(s): Cheng, T., Hitomi K., Schalkwijk, J. Zeeuwen, P.L.J.M., (他3名)
  • Journal Title: Br J. Dermatol. Volume: 161 Pages: 253-264
  • Peer Reviewed
• [Journal Article] タンパク質架橋化酵素の高反応性基質配列の探索と活用 (2009)
  • Author(s): 人見清隆
  • Journal Title: 生化学 Volume: vol81.No.8. Pages: 708-711
  • NAID: 10026298303
  • Peer Reviewed
• [Journal Article] Development of an isozyme-specific colorimetric assay for tissue transglutaminase 2 cross-linking assay (2009)
  • Author(s): Alea, M. P., Kitamura, M., Martin, G., Thomas, V., Hitomi, K., Alaoui, S El.
  • Journal Title: Anal. Biochem. Volume: 389 Pages: 150-156
  • Peer Reviewed
• [Journal Article] A Specific Colorimetric assay for measuring of transglutaminase 1 and factor XIII activities. (2009)
  • Author(s): Hitomi, K., Kitamura, M., Alea, M.P., Ismail, Thomas, V., Alaoui. S El.
  • Journal Title: Analytical Biochemistry 394 Pages: 281-284
  • Peer Reviewed
• [Journal Article] The cystatin M/E-controlled of skin barrier formation : expression of its key components in psoriasis and atopic dermatitis. (2009)
  • Author(s): Cheng, T., Tiabringa GS, van-Vlijmen-Willems IM, Hitomi K, van Erp PE, Shalkwilk J, Zeeuwen, PM
  • Journal Title: British Journal of Dermatology 161 Pages: 253-264
  • Peer Reviewed
• [Journal Article] Preferred substrate sequences for transglutaminase 2 : screening using a phage-displayed peptide library (2009)
  • Author(s): Hitomi K., Kitamura Y.,Sugimura Y
  • Journal Title: Amino Acids 36 Pages: 619-624
  • Peer Reviewed
• [Journal Article] タンパク質架橋化酵素の高反応性基質配列の探索と活用 (2009)
  • Author(s): 人見清隆
  • Journal Title: 生化学 8 Pages: 708-711
  • NAID: 10026298303
• [Journal Article] Development of an isozyme-specific colorimetric assay for tissue transglutaminase 2 cross-linking assay. (2009)
  • Author(s): Alea, M.P., Kitamura, M., Martin, G., Thomas, V., Hitomi, K., Alaoui, S El.
  • Journal Title: Analytical Biochemistry 389 Pages: 150-156
  • Peer Reviewed
• [Presentation] トランスグルタミナーゼが触媒するタンパク質のポリアミン修飾による生理機能の調節 (2012)
  • Author(s): 人見清隆
  • Organizer: 日本農芸化学会(大会シンポジウム)
  • Place of Presentation: 京都女子大学(招待講演)
  • Year and Date: 2012-03-25
• [Presentation] ヒト前立腺型タンパク質架橋化酵素の発現と前立腺由来タンパク質の基質配列解析 (2012)
  • Author(s): 樋口幾、福井美奈、伊藤みほ、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 2012-03-24
• [Presentation] モデル生物としてのメダカ(Oryzias latipes)におけるタンパク質架橋化酵素の解析 (2012)
  • Author(s): 斉藤麻衣、樋口幾、福井美奈、伊藤みほ、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 2012-03-24
• [Presentation] Characterization of medaka transglutaminases, a protein cross-linking enzyme family (2011)
  • Author(s): 斉藤麻衣、伊藤みほ、人見清隆
  • Organizer: Startegic Meeting for Medaka Reasearch
  • Place of Presentation: Okazaki Conference Center, Aichi(招待講演)
  • Year and Date: 2011-11-23
• [Presentation] タンパク質架橋化酵素トランスグルタミナーゼの高反応性基質配列の探索と多面的活用 (2011)
  • Author(s): 人見清隆
  • Organizer: 酵素工学研究会
  • Place of Presentation: 東京大学山上会館(招待講演)
  • Year and Date: 2011-09-29
• [Presentation] 蛋白質架橋化酵素ファミリーの新規アイソザイムの発現解析 (2011)
  • Author(s): 清水由隆、福井美奈、伊藤みほ、人見清隆
  • Organizer: 日本生化学会大会
  • Place of Presentation: 京都国際会議場(招待講演)
  • Year and Date: 2011-09-23
• [Presentation] 新規なヒト蛋白質架橋化酵素アイソザイムの高反応性基質配列の解析 (2011)
  • Author(s): 鞍本克真、福井美奈、伊藤みほ、人見清隆
  • Organizer: 日本生化学会大会
  • Place of Presentation: 京都国際会議場
  • Year and Date: 2011-09-23
• [Presentation] タンパク質架橋化酵素の蛍光標識高反応性基質ペプチドを用いたマウス胎児組織 (2011)
  • Author(s): 伊藤みほ、川本忠文、人見清隆
  • Organizer: 日本生化学会大会
  • Place of Presentation: 京都国際会議場
  • Year and Date: 2011-09-23
• [Presentation] Identification of the preferred substrate sequences for transglutaminase and utility for detection of activity in an isozyme-specific manner (2011)
  • Author(s): 人見清隆
  • Organizer: 日本生化学会大会シンポジウム
  • Place of Presentation: 京都国際会議場(招待講演)
  • Year and Date: 2011-09-21
• [Presentation] タンパク質架橋化酵素の高反応性基質配列の探索と活用 (2011)
  • Author(s): 伊藤みほ、福井美奈、鞍本克真、清水由隆、人見清隆
  • Organizer: 動物細胞工学会
  • Place of Presentation: 東京大学山上会館
  • Year and Date: 2011-07-23
• [Presentation] 全マウス組織切片を用いたタンパク質架橋化酵素活性の可視化 (2011)
  • Author(s): 伊藤みほ、川本忠文、柴田秀樹、牧正敏、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: (要旨集)
  • Year and Date: 2011-03-27
• [Presentation] 新規タンパク質架橋化酵素Tgase 6の高反応性基質配列の探索 (2011)
  • Author(s): 鞍本克真、福井美奈、清水由隆、柴田秀樹、牧正敏、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: (要旨集)
  • Year and Date: 2011-03-27
• [Presentation] Characterization of medaka transg/Utamirlases, a protein cross-linking enzyme family (2011)
  • Author(s): 斉藤麻衣、樋口幾、伊藤みほ、人見清隆
  • Organizer: Medaka Strategic Research
  • Place of Presentation: 岡崎(愛知県)
• [Presentation] タンパク質架橋化酵素トランスグルタミナーゼの高反応性基質配列の探索と多面的活用 (2011)
  • Author(s): 人見清隆
  • Organizer: 酵素工学会
  • Place of Presentation: 東京(東京大学山上会館)
• [Presentation] Identification of the preferred substrate sequences for transglutaminase and utility for detection of activity in an isozyme-specific manner (2011)
  • Author(s): 人見清隆
  • Organizer: 日本生化学会大会シンポジウム
  • Place of Presentation: 京都国際会議場
• [Presentation] タンパク質架橋化酵素の高反応性基質配列の探索と活用 (2011)
  • Author(s): 伊藤みほ、福井美奈、鞍本克真、清水由隆、人見清隆
  • Organizer: 日本動物細胞工学会
  • Place of Presentation: 東京(東京大学山上会館)
• [Presentation] Identification of highly reactive substrate peptides for transglutaminases from random peptide library and their applications (2010)
  • Author(s): Kiyotaka Hitomi
  • Organizer: Gordon Research Conference
  • Place of Presentation: Davidson College(米国)
  • Year and Date: 2010-07-19
• [Presentation] 高等動物トランスグルタミナーゼの高反応性基質配列の探索と活用 (2010)
  • Author(s): 人見清隆
  • Organizer: 日本農芸化学会2010年大会
  • Place of Presentation: 東京大学(東京都)
  • Year and Date: 2010-03-30
• [Presentation] 機能未知なタンパク質架橋化酵素(TG6)の高反応性基質配列の探索 (2010)
  • Author(s): 福井美奈、山根亜沙佳、伊藤みほ、人見清隆
  • Organizer: 生化学会大会分子生物学会年会
  • Place of Presentation: 神戸国際会議場
• [Presentation] 表皮特異的タンパク質架橋化酵素(TG3)の高反応性基質配列の解析 (2010)
  • Author(s): 山根亜沙佳、福井美奈、伊藤みほ、人見清隆
  • Organizer: 生化学会大会分子生物学会年会
  • Place of Presentation: 神戸国際会議場
• [Presentation] 分化段階の骨芽細胞株におけるタンパク質架橋化酵素トランスグルタミナーゼの基質解析 (2010)
  • Author(s): 渡邊一哉、人見清隆
  • Organizer: 生化学会大会分子生物学会年会
  • Place of Presentation: 神戸国際会議場
• [Presentation] タンパク質架橋化酵素の高反応性基質ペプチドを活用したマウス全組織内酵素活性の可視化 (2010)
  • Author(s): 伊藤みほ、川本忠文、人見清隆
  • Organizer: 農芸化学会中部支部例会
  • Place of Presentation: 名古屋(名古屋大学)
• [Presentation] Screeninig and identification of the preferred substrate sequences for tranglutaminase 3 using phage-display peptide library (2010)
  • Author(s): 山根亜沙佳、福井美奈、伊藤みほ、人見清隆
  • Organizer: ゴードン国際会議
  • Place of Presentation: 米国
• [Presentation] Detection of in situ activities of transglutaminases using highly reactive substrate peptides (2010)
  • Author(s): 伊藤みほ、川本忠文、人見清隆
  • Organizer: ゴードン国際会議
  • Place of Presentation: 米国
• [Presentation] Identification of highly reactive substrate peptides for transglutaminases from random peptide library and its application (2010)
  • Author(s): 人見清隆
  • Organizer: ゴードン国際会議
  • Place of Presentation: 米国
• [Presentation] タンパク質架橋化酵素トランスグルタミナーゼの高反応性基質配列の解明と活用 (2010)
  • Author(s): 人見清隆
  • Organizer: 天野エンザイム応用酵素シンポジウム
  • Place of Presentation: 名古屋
• [Presentation] Establishment of a novel assay system for transglutaminase using highly reactive substrate peptide (2010)
  • Author(s): 人見清隆
  • Organizer: 国際ポリアミン学会
  • Place of Presentation: 静岡(御殿場)
• [Presentation] ヒト表皮型タンパク質架橋化酵素(TGase 3)の高反応性基質配列の探索と解析 (2010)
  • Author(s): 福井美奈、山根亜沙佳、伊藤みほ、人見清隆, 他
  • Organizer: 生化学会中部支部例会
  • Place of Presentation: 名古屋(名古屋大学)
• [Presentation] ヒト表皮型トランスグルタミナーゼに対する高反応性基質配列の探索と解析 (2010)
  • Author(s): 福井美奈、山根亜沙佳、柴田秀樹、牧正敏、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 東京・東京大学
• [Presentation] 高等動物トランスグルタミナーゼの高反応性基質配列の探索と活用 (2010)
  • Author(s): 人見清隆
  • Organizer: 日本農芸化学会大会シンポジウム
  • Place of Presentation: 東京・東京大学
• [Presentation] タンパク質を架橋接着する酵素(トランスグルタミナーゼ)の高反応性基質配列の探索と多面的活用 (2009)
  • Author(s): 人見清隆
  • Organizer: 山形バイオカフェ
  • Place of Presentation: 山形県鶴岡市先端研究産業センター(山形県)
  • Year and Date: 2009-12-16
• [Presentation] タンパク質架橋酵素の高反応性基質ペプチドの探索とその有効活用 (2009)
  • Author(s): 人見清隆
  • Organizer: 日本生化学会第82回大会
  • Place of Presentation: 神戸国際会議場(神戸市)
  • Year and Date: 2009-10-23
• [Presentation] 生体内タンパク質接着酵素反応における高反応性基質ペプチドの探索とその多面的活用 (2009)
  • Author(s): 人見清隆
  • Organizer: 東京慈恵会医科大学セミナー
  • Place of Presentation: 慈恵会医科大学(東京都)
  • Year and Date: 2009-09-11
• [Presentation] Characterization of the preferred substrate sequence for transglut aminase, an enzyme for protein cross-linking and polyamine-conjugation (2009)
  • Author(s): Kiyotaka Hitomi
  • Organizer: Gordon Research Conference : Polyamines
  • Place of Presentation: Waterville Valley (NJ, USA)
  • Year and Date: 2009-06-23
• [Presentation] トランスグルタミナーゼの骨芽細胞株における発現と架橋化酵素活性の解析 (2009)
  • Author(s): 渡辺一哉、角田佳奈子、柴田秀樹、牧正敏、人見清隆
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸国際展示場
• [Presentation] タンパク質架橋酵素の高反応性基質ペプチドの探索とその有効活用 (2009)
  • Author(s): 人見清隆
  • Organizer: 第82回日本生化学会大会シンポジウム
  • Place of Presentation: 神戸国際会議場
• [Presentation] 生体内タンパク質接着反応における高反応性基質と多面的活用 (2009)
  • Author(s): 人見清隆
  • Organizer: 慈恵会医科大学分子生物セミナー
  • Place of Presentation: 東京・慈恵会医科大学
• [Presentation] (2009)
  • Author(s): Kiyotaka HITOMI
  • Organizer: ゴードン国際会議「ポリアミン」
  • Place of Presentation: アメリカニューハンプシャー州
• [Presentation] トランスグルタミナーゼに対する高反応性基質ペプチドを活用した活性検出法の確立
  • Author(s): 伊藤みほ、柴田秀樹、牧正敏、人見清隆
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 東京・東京大学