Research activity information

• Rapid inactivation of the yeast Sec complex selectively blocks transport of post-translationally translocated proteins

  Jae Kyo Yi, Hidenobu Fujita, Elisabet C. Mandon, Reid Gilmore

  Elsevier BV, Oct. 2021, Journal of Biological Chemistry, 297(4) (4), 101171 - 101171

  Scientific journal


• Afadin Facilitates Vascular Endothelial Growth Factor-Induced Network Formation and Migration of Vascular Endothelial Cells by Inactivating Rho-Associated Kinase Through ArhGAP29.

  Toru Tagashira, Terunobu Fukuda, Muneaki Miyata, Kazuha Nakamura, Hidenobu Fujita, Yoshimi Takai, Ken-Ichi Hirata, Yoshiyuki Rikitake

  OBJECTIVE: We previously reported that afadin, an actin filament-binding protein, regulated vascular endothelial growth factor-induced angiogenesis. However, the underlying molecular mechanisms are poorly understood. Here, we investigated the mechanisms of how Rho-associated kinase is activated in afadin-knockdown human umbilical vein endothelial cells (HUVECs) and how its activation is involved in defects of vascular endothelial growth factor-induced network formation and migration of the cells. APPROACH AND RESULTS: Knockdown of afadin or ArhGAP29, a GTPase-activating protein for RhoA, increased Rho-associated kinase activity and reduced the vascular endothelial growth factor-induced network formation and migration of cultured HUVECs, accompanied by the defective formation of membrane protrusions, such as lamellipodia and peripheral ruffles. Treatment of the afadin- or ArhGAP29-knockdown HUVECs with Rho-associated kinase inhibitors, Y-27632 or fasudil, partially restored the reduced network formation and migration as well as the defective formation of membrane protrusions. ArhGAP29 bound to afadin and was colocalized with afadin at the leading edge of migrating HUVECs. The defective formation of membrane protrusions in ArhGAP29-knockdown HUVECs was restored by expression of mutant ArhGAP29 that bound to afadin and contained a RhoGAP domain but not mutant ArhGAP29 that could bind to afadin and lacked the RhoGAP domain or mutant ArhGAP29 that could not bind to afadin and contained the RhoGAP domain. This suggested the requirement of both the interaction of afadin with ArhGAP29 and RhoGAP activity of ArhGAP29 for migration of HUVECs. CONCLUSIONS: Our results highlight a critical role of the afadin-ArhGAP29 axis for the regulation of Rho-associated kinase activity during vascular endothelial growth factor-induced network formation and migration of HUVECs.

  May 2018, Arteriosclerosis, thrombosis, and vascular biology, 38(5) (5), 1159 - 1169, English, International magazine

  Scientific journal


• Necl-4 enhances the PLCγ-c-Raf-MEK-ERK pathway without affecting internalization of VEGFR2.

  Shota Yamana, Amina Tokiyama, Hidenobu Fujita, Yuya Terao, Sayo Horibe, Naoto Sasaki, Seimi Satomi-Kobayashi, Ken-Ichi Hirata, Yoshiyuki Rikitake

  We have reported that knockdown of Necl-4 decreases vascular endothelial growth factor (VEGF)-induced phosphorylation of extracellular signal-regulated kinase (ERK) without affecting phosphorylation of VEGF receptor 2 (VEGFR2) in sparsely cultured human umbilical vein endothelial cells (HUVECs). However, the underlying molecular mechanism is unknown. Compared with control HUVECs, VEGF-induced phosphorylation of phospholipase Cγ (PLCγ), c-Raf, mitogen-activated protein kinase/ERK kinase (MEK) and ERK were all inhibited in Necl-4-knockdown HUVECs. However, VEGF-induced internalization of VEGFR2 was not different between control and Necl-4-knockdown HUVECs. We have reported that protein-tyrosine phosphatase, non-receptor type 13 (PTPN13) and Rho-associated kinase (ROCK) are involved in the Necl-4-knockdown-induced inhibition of the VEGF-induced activation of Rac1. However, the effects of Necl-4-knockdown on VEGF-induced phosphorylation of VEGFR2 and ERK were not affected either by knockdown of PTPN13 or by ROCK inhibitors. These results suggest that Necl-4 enhances VEGF-induced activation of PLCγ-c-Raf-MEK-ERK pathway without affecting the phosphorylation and internalization of VEGFR2.

  Aug. 2017, Biochemical and biophysical research communications, 490(2) (2), 169 - 175, English, International magazine

  Scientific journal


• Interaction of FAM5C with UDP-glucose:glycoprotein glucosyltransferase 1 (UGGT1): Implication of N-glycosylation in FAM5C secretion

  Yuya Terao, Hidenobu Fujita, Sayo Horibe, Junya Sato, Satomi Minami, Miwako Kobayashi, Ichiro Matsuoka, Naoto Sasaki, Seimi Satomi-Kobayashi, Ken-ichi Hirata, Yoshiyuki Rikitake

  May 2017, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 486(3) (3), 811 - 816, English

  [Refereed]

  Scientific journal


• Stability and flexibility of marginally hydrophobic-segment stalling at the endoplasmic reticulum translocon

  Yuichiro Kida, Yudai Ishihara, Hidenobu Fujita, Yukiko Onishi, Masao Sakaguchi

  Mar. 2016, MOLECULAR BIOLOGY OF THE CELL, 27(6) (6), 930 - 940, English

  [Refereed]

  Scientific journal


• A Few Positively Charged Residues Slow Movement of a Polypeptide Chain across the Endoplasmic Reticulum Membrane

  Marifu Yamagishi, Yukiko Onishi, Shotaro Yoshimura, Hidenobu Fujita, Kenta Imai, Yuichiro Kida, Masao Sakaguchi

  Aug. 2014, BIOCHEMISTRY, 53(33) (33), 5375 - 5383, English

  [Refereed]

  Scientific journal


• Stop-and-Move of a Marginally Hydrophobic Segment Translocating across the Endoplasmic Reticulum Membrane

  Yukiko Onishi, Marifu Yamagishi, Kenta Imai, Hidenobu Fujita, Yuichiro Kida, Masao Sakaguchi

  Sep. 2013, JOURNAL OF MOLECULAR BIOLOGY, 425(17) (17), 3205 - 3216, English

  [Refereed]

  Scientific journal


• Pleiotropic Effects of Membrane Cholesterol upon Translocation of Protein across the Endoplasmic Reticulum Membrane

  Hitoshi Yamamoto, Hidenobu Fujita, Yuichiro Kida, Masao Sakaguchi

  May 2012, BIOCHEMISTRY, 51(17) (17), 3596 - 3605, English

  [Refereed]

  Scientific journal


• Positive charges on the translocating polypeptide chain arrest movement through the translocon

  Hidenobu Fujita, Marifu Yamagishi, Yuichiro Kida, Masao Sakaguchi

  Dec. 2011, JOURNAL OF CELL SCIENCE, 124(24) (24), 4184 - 4193, English

  [Refereed]

  Scientific journal


• A sugar chain at a specific position in the nascent polypeptide chain induces forward movement during translocation through the translocon

  Marifu Yamagishi, Hidenobu Fujita, Fumiko Morimoto, Yuichiro Kida, Masao Sakaguchi

  May 2011, JOURNAL OF BIOCHEMISTRY, 149(5) (5), 591 - 600, English

  [Refereed]

  Scientific journal


• Positive Charges of Translocating Polypeptide Chain Retrieve an Upstream Marginal Hydrophobic Segment from the Endoplasmic Reticulum Lumen to the Translocon

  Hidenobu Fujita, Yuichiro Kida, Masatoshi Hagiwara, Fumiko Morimoto, Masao Sakaguchi

  Jun. 2010, MOLECULAR BIOLOGY OF THE CELL, 21(12) (12), 2045 - 2056, English

  [Refereed]

  Scientific journal

• the mechanism of membrane protein folding through the translocon

  藤田英伸, 阪口雅郎

  2022, 日本分子生物学会年会プログラム・要旨集(Web), 45th


• 小胞体トランスロコンでの膜タンパク質の構造形成における膜貫通セグメント識別の2つの容態

  阪口雅郎, 高原教代, 藤田英伸

  2018, 日本蛋白質科学会年会プログラム・要旨集, 18th


• 小胞体トランスロコンでのポリペプチド鎖の動き制限要因と駆動要因

  藤田英伸, 高原教代, 木田祐一郎, 阪口雅郎

  2018, 日本生化学会大会(Web), 91st


• Importance of N-glycosylation in the Secretion of FAM5C interecting with UDP-glucose glycoprotein glycosyltransferase (UGGT1)

  寺尾侑也, 寺尾侑也, 藤田英伸, 堀部紗世, 佐藤淳哉, 佐藤淳哉, 南智美, 平田健一, 力武良行, 力武良行, 力武良行

  2016, 日本生化学会大会(Web), 89th


• ラミニンによるアストロサイト突起形成の促進

  藤田英伸, 佐藤淳哉, 堀部紗世, 南智美, 平田健一, 力武良行, 力武良行

  2016, 日本分子生物学会年会プログラム・要旨集(Web), 39th


• 小胞体を介したタンパク質膜透過におけるSec62/63複合体の役割

  藤田英伸, MANDON Elisabet, GILMORE Reid

  2013, 日本分子生物学会年会プログラム・要旨集(Web), 36th


• 粗面小胞体上のリボソームが膜貫通配列を認識する

  木田祐一郎, 石原裕大, 藤田英伸, 阪口雅郎

  2013, 日本細胞生物学会大会要旨集, 65th


• 小胞体トランスロコンを介したポリペプチド鎖の動き

  藤田英伸, 木田祐一郎, 阪口雅郎

  2012, 日本蛋白質科学会年会プログラム・要旨集, 12th


• Effects of positive charges on the movement of polypeptide chain through the ER translocon

  FUJITA Hidenobu, KIDA Yuichiro, YAMAGISHI Marifu, SAKAGUCHI Masao

  2011, 生物物理, 51(Supplement 1) (Supplement 1)


• ポリペプチド鎖の環境変化が示すトランスロコン通過挙動

  木田祐一郎, 藤田英伸, 久米千智, 平野真希, 阪口雅郎

  2010, 日本生体エネルギー研究会討論会講演要旨集, 36th

• THE MOLECULAR BIOLOGY SOCIETY OF JAPAN


• PROTEIN SCIENCE SOCIETY OF JAPAN


• THE JAPANESE BIOCHEMICAL SOCIETY

• 小胞体トランスロコンを介した膜タンパク質形成時のタンパク質の動態の解明

  藤田 英伸

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 兵庫県立大学, 01 Apr. 2022 - 31 Mar. 2025


• 小胞体膜トランスロコンにおけるタンパク質膜透過の動態

  藤田英伸

  公益財団法人 ひょうご科学技術協会, 兵庫県特別研究 助成金, 兵庫県立大学大学院 理学研究科, Apr. 2021 - Mar. 2022, Principal investigator

  Competitive research funding


• 小胞体トランスロコンによる膜タンパク質構 造形成機構の解明

  藤田英伸

  公益財団法人 ひょうご科学技術協会, 兵庫県特別研究 助成金, 兵庫県立大学大学院 理学研究科, Apr. 2019 - Mar. 2020