The function of ubiquitin in intralysosomal protein degradation.

• Project/Area Number: 16590235
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Tokushima Bunri University
• Principal Investigator: ISHIDOH Kazumi Tokushima Bunri University, Intsitute for Health Sciences, Division of Molecular Biology, Professor, 健康科学研究所・分子生物学部門, 教授 (40212906)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: lysosome / protein degradation / ubiquitin / insulin receptor / epidermal growth factor receptor / platelet derived growth factor receptor / LDH-A / ユビキチン / ポリユビキチン化 / マルチユビキチン化 / c-Cbl

Research Abstract

HA-c-Cbl was expressed in HEK293 cells and immunoprecipitated. Insulin receptor expressed in HIRc cells was immunoprecipitaed. Then Both immunoprecipitants were mixed with E1,UbcH7 as an E2,ATP and its regenerating system, ubiquitin or its mutants. These mixtures were reacted in pH6.0, 6.5 and 7.2. Ubiquitinated insulin receptor were detected by immunoblot. When the reaction was done at pH7.2, insulin receptor were monoubiquitinated with multiple lysine residues in insulin receptor, whereas when the reaction was carried out at pH6.0, the insulin receptor was subjected to polyubiqutination through Lys-48 in ubiquitin molecules. Therefore, I projected next to analyze the structural change of recombinant c-Cbl between neutral pH and acidic pH.
Next, I was planed to analyze the ubiquitination profiles of epidermal growth factor receptor and platelet derived growth factor receptor. These projects would be under going.
Recently, I identified the monoubiquitination of LDH-A, a cytosolic protein, under oxidative stress was sequestered into and degraded in lysosomes. Now, I would search the ubiquitin receptor on the cytosolic surface of lysosomal membranes.

Research Products

• [Journal Article] CDK5 regulates cell0cell and cell-matrix adhesion in human keratinocytes. (2005)
  • Author(s): Nakano N
  • Journal Title: Br J Dermatol 153・1 Pages: 37-45
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of mono-ubiquitinated LDH-A in skeletal muscle cells exposed to oxidative stress. (2005)
  • Author(s): Onishi Y
  • Journal Title: Biochem Biophys Res Commun 336・3 Pages: 799-806
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells (2005)
  • Author(s): Hashimoto M
  • Journal Title: Int J Parasitol 25・14 Pages: 1587-1594
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ubiquitin ligase Cbl-b downregukates bone formation through suppression of IGF-I signaling in osteoblasts during denervation (2005)
  • Author(s): Suzue N
  • Journal Title: J Bone Mineral Res (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Microarrayanalysis identified differentially expressed genes in keratocytes from keratocounus patients. (2005)
  • Author(s): Ha N.T., Nakayasu, K., Murakami, A., Ishidoh K., Kanai A.
  • Journal Title: Curr.Eye Res. 28 Pages: 373-379
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] CDK5 regulates cell-cell and cell-matrix adhesion in human keratinocytes. (2005)
  • Author(s): Nakano N, Nakao, A.Ishidoh, K., Kominami, E., Tsuboi, R., Ogawa, H.
  • Journal Title: Br.J.Dermatol. 153 Pages: 37-45
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of mono-ubiquitinated LDH-A in skeletal muscle cells exposed to oxidative stress. (2005)
  • Author(s): Onishi Y, Hirasaka K, Ishihara I, Oarada M, Goto J, Ogawa T, Suzue N, Nakano S, Furochi H, Ishidoh K, Kishi K, Nikawa T.
  • Journal Title: Biochem.Biophys.Res.Commun. 336 Pages: 799-806
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells. (2005)
  • Author(s): Hashimoto M, Nakajima-Shimada J, Ishidoh K, Aoki T.
  • Journal Title: Int.J.Parasitol. 25 Pages: 1587-1594
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells. (2005)
  • Author(s): Hashimoto M
  • Journal Title: Int J Parasitol. 35・14 Pages: 1587-1594
• [Journal Article] Identification of mono-ubiquitinated LDH-A in skeltalmuscle exposed to oxidative stress. (2005)
  • Author(s): Onishi Y
  • Journal Title: Biochem.Biophys.Res.Commun. 336・3 Pages: 799-806
• [Journal Article] CDK5 regulates cell-cell and cell-matrix adhesion in human keratinocytes. (2005)
  • Author(s): Nakano N
  • Journal Title: Br.J.Dermatol. 153・1 Pages: 37-45
• [Journal Article] Skeltal muscle gene expression in space-flown rats. (2004)
  • Author(s): Nikawa T
  • Journal Title: FASEB J 18・3 Pages: 522-524
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inflammatory response and cathepsins in silica-induced ahermansky-Pudlock syndrome model pale ear mice/ (2004)
  • Author(s): Yoshioka Y
  • Journal Title: Pathol Int 54・5 Pages: 322-331
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mammalian class E Vps proteins, SBP1 and mVps/CHMP2A, interact and regulate the function of an AAA-ATP汗SKD1/Vps4B (2004)
  • Author(s): Fujita H
  • Journal Title: J Cell Sci 117・4 Pages: 2997-3009
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Podocyte migration during nephrotic syndrome requires a coordinated interplay between cathepsin L and α3 integurin. (2004)
  • Author(s): Reiser J
  • Journal Title: J Biol Chem 279・33 Pages: 34827-34832
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Proteinases participating in the processing and activation of prolegumain in cultured primary macrophages. (2004)
  • Author(s): Lecaille F
  • Journal Title: Biol Chem 385・6 Pages: 511-515
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Epidermal growth factor activates m-calpain, resulting in apoptosis of HaCaT keratinocytes. (2004)
  • Author(s): Inoue A
  • Journal Title: J Dermatol Sci 36・1 Pages: 60-62
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Microarray analysis identified differentially expressed genes in keratocytes from keratoconus patients. (2004)
  • Author(s): Na NT
  • Journal Title: Curr Eye Res. 28・6 Pages: 373-379
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 糖尿病とタンパク質のユビキチン化 (2004)
  • Author(s): 石堂一巳
  • Journal Title: 内分泌・糖尿病化 18・4 Pages: 352-357
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Skeltal muscle gene expression in space-flown rats. (2004)
  • Author(s): Nikawa, T., Ishidoh, K., Hirasaka, K., Ishihara, K., Ikemoto, M., Kano, K., Kominami, E., NOnaka, I., Ogawa, T., Adam, G.R., Baldwin, K.M., Yasui, N., Kishi, K., Takeda, S.
  • Journal Title: FASEB.J. 18 Pages: 522-524
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inflammatory response and cathepsins in silica-exposed Hermansky-Pudlock syndrome medel pale year mice. (2004)
  • Author(s): Yoshioka, Y., Kumasaka, T., Ishidoh, K., Kominami, E., Mitani, K., Hosokawa, Y., Fukuchi, Y.
  • Journal Title: Pathology Int. 54 Pages: 322-331
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mammalian class E Vps proteins, SBP1 and mVps2/CHMP2A, interact with and regulate the function of an AAA-ATPase SKD1/Vps4B. (2004)
  • Author(s): Fujita, H., Umezaki, Y., Imamura, K., Ishikawa, D., Uchimura, S., Nara, A., Yoshimori, T., Hayashizaki, Y., Kawai, J., Ishidoh, K., Tanaka Y., Himeno, M.
  • Journal Title: J.Cell Sci. 117 Pages: 2997-3009
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Epidermal growth factor activates m-calpain, resulting in apoptosis in HaCaT keratinocytes. (2004)
  • Author(s): Inoue, A., Yamazaki, M., Ishidoh, K., Ogawa, H.
  • Journal Title: J.Dermatol Sci. 36 Pages: 60-62
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Analysis on which types of proteinases participate in processing and activation of prolegumain in primary cultured rat macrophages. (2004)
  • Author(s): Lecaille, F., Muno, D., Kominami, E., Ishidoh, K.
  • Journal Title: Biol.Chem. 385 Pages: 511-516
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Podocyte migration during nephrotic syndrome requires a coordinated interplay between cathepsin L and a3 integrin. (2004)
  • Author(s): Reiser J, Oh J, Shirato I, Asanuma K, Hug A, Mundel TM, Honey K, Ishidoh K, Kominami E, Kreidberg JA, Tomino Y, Mundel P.
  • Journal Title: J.Biol.Chem. 279 Pages: 34827-34832
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Microarray analysis identified differentially expressed genes in keratocytes from keratocounus patients. (2004)
  • Author(s): Ha, NT, Nakayasu, K, Murakami, A, Ishidoh, K, Kanai, A
  • Journal Title: Curr.Eye Research 28 Pages: 373-379
• [Journal Article] Podocyte migration during nephrotic syndrome requires a coordinated interplay between cathepsin L and α3 integrin. (2004)
  • Author(s): Reiser, J, Oh, J, Shirato, I, Asanuma, K, Hug, A, Mundel, TM, (他6名)
  • Journal Title: J Biol Chem 279 Pages: 34827-34832
• [Journal Article] Analysis on which types of proteinases participate in processing and activation of prolegumain in primary cultured rat macrophages. (2004)
  • Author(s): Lecaille, F, Muno, D, Kominami, E., Ishidoh K
  • Journal Title: Biol Chem 385 Pages: 511-516
• [Journal Article] Mammalian calss E Vps proteins, SBP1 and mVPS2/CHMP2A, interact with and regulate the function of an AAA-ATPase SKD1/Vps4B. (2004)
  • Author(s): Fujita, H, Umezaki, Y, Imamura, K, (他9名)
  • Journal Title: J Cell Sci 117 Pages: 2997-3009
• [Journal Article] Skeltal muscle gene expression in space-flown rats. (2004)
  • Author(s): NIkawa, T, Ishidoh, K, Hirasaka, K, Ishihara, K, (他10名)
  • Journal Title: FASEB J 18 Pages: 522-524
• [Journal Article] Ubiquitin ligase Cbl-b downregulates bone formation through suppression of IGF-I signaling in osteoblasts during denervation.
  • Author(s): Suzue N, Nikawa T, Onishi Y, Yamada, C, Hirasaka K, Ogawa T, Furochi H, Kosaka H, Ishidoh K, Gu H, Takeda S, Ishimaru N, Hayashi Y, Yamamoto H, Kishi K, Yasui N
  • Journal Title: J.Bone Mineral Res. (In press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Ubiquitin ligase Cb1-b downregulates bone formation through suppression of IGF-I signaling in osteroblast during denervation.
  • Author(s): Suzue N
  • Journal Title: J Bone Meneral Res (In press)
• [Journal Article] CDK5 regulates cell-cell and cell-matrix adhesion in human keratinocytes
  • Author(s): Nakano, N, Nakao, A, Murakami, A, Ishidoh K, (他3名)
  • Journal Title: Br.J.Dermatol in press
• [Book] 癌転移とプロテアーゼinプロセテーゼインヒビター臨床編 (2005)
  • Author(s): 石堂一巳
  • Total Pages: 271
  • Publisher: 診断と治療社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 癌転移とプロテアーゼ プロテアーゼインヒビター臨床編 (2005)
  • Author(s): 石堂一巳
  • Total Pages: 271