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Inhibition of the EGF-induced activation of phospholipase C-γ1 by a single chain antibody fragment

Oncogene volume 20, pages 7954–7964 (2001)Cite this article

Abstract

Phospholipase C-γ1(PLC-γ1) is known to play an essential role in various cellular responses, such as proliferation and tumorigenesis, and PLC-γ1-specific inhibitors are commonly employed to investigate the mechanism of the PLC-γ1-mediated signaling pathway. In this study, we developed a single chain antibody fragment (scFv) as a blocker for PLC-γ1 mediated signaling. scFv, designated F7-scFv, specifically bound to PLC-γ1 with high affinity (Kd=1.9×10−8 M) in vitro. F7-scFv also bound to PLC-γ1 in vivo and altered the distribution pattern of PLC-γ1 from the cytoplasm to the intracellular aggregates, where F7-scFv was localized. Moreover, F7-scFv interrupted the EGF-induced translocation of PLC-γ1 from the cytosol to the membrane ruffle and attenuated EGF-induced inositol phosphates generation and intracellular calcium mobilization. These results indicate that F7-scFv blocks EGF-induced PLC-γ1 activation by causing sequestering of PLC-γ1 into intracellular aggregates, and may therefore be useful in studies of the PLC-γ1-mediated signaling pathway.

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References

  • Anderson D, Koch CA, Grey L, Ellis C, Moran MF, Pawson T . 1990 Science 250: 979–982

  • Arteaga CL, Johnson MD, Todderud G, Coffey RJ, Carpenter G, Page DL . 1991 Proc. Natl. Acad. Sci. USA 88: 10435–10439

  • Bae SS, Perry DK, Oh YS, Choi JH, Galadari SH, Ghayur T, Ryu SH, Hannun YA, Suh PG . 2000 FASEB J. 14: 1083–1092

  • Beerli RR, Wels W, Hynes NE . 1994 J. Biol. Chem. 269: 23931–23936

  • Berridge MJ . 1993 Nature 361: 315–325

  • Biocca S, Cattaneo A . 1995 Trends Cell. Biol. 5: 248–252

  • Cardinale A, Lener M, Messina S, Cattaneo A, Biocca S . 1998 FEBS Lett. 439: 197–202

  • Cattaneo A, Biocca S . 1999 Trends Biotechnol. 17: 115–121

  • Chang JS, Noh DY, Park IA, Kim MJ, Song H, Ryu SH, Suh PG . 1997 Cancer Res. 57: 5465–5468

  • Cochet O, Kenigsberg M, Delumeau I, Virone-Oddos A, Multon MC, Fridman WH, Schweighoffer F, Teillaud JL, Tocque B . 1998 Cancer Res. 58: 1170–1176

  • Cockcroft S, Thomas GM . 1992 Biochem. J. 288: 1–14

  • Diakonova M, Payrastre B, van Velzen AG, Hage WJ, van Bergen en Henegouwen PM, Boonstra J, Cremers FF, Humbel BM . 1995 J. Cell. Sci. 108: 2499–2509

  • Hoogenboom HR, Bruine AP, Hufton SE, Heot RM, Arends JW, Roovers RC . 1998 Immunotechnology 4: 1–20

  • Huang PS, Davis L, Huber H, Goodhart PJ, Wegrzyn RE, Oliff A, Heimbrook DC . 1995 FEBS Lett. 358: 287–292

  • Ji QS, Winnier GE, Niswender KD, Horstman D, Wisdom R, Magnuson MA, Carpenter G . 1997 Proc. Natl. Acad. Sci. USA 94: 2999–3003

  • Kabat EA, Wu TT, Perry HM, Gottesman KS, Foeller C . 1991 Sequences of proteins of immunological interest, 5th ed National Institutes of Health, Bethesda, MD

  • Katan M . 1998 Biochim. Biophys. Acta 1436: 5–17

  • Kim HK, Kim JW, Zilberstein A, Margolis B, Kim JG, Schlessinger J, Rhee SG . 1991 Cell 65: 435–441

  • Kim JW, Sim SS, Kim UH, Nishibe S, Wahl MI, Carpenter G, Rhee SG . 1990 J. Biol. Chem. 265: 3940–3943

  • Kim MJ, Chang JS, Park SK, Hwang JI, Ryu SH, Suh PG . 2000 Biochemistry 39: 8674–8682

  • Koch CA, Anderson D, Moran MF, Ellis C, Pawson T . 1991 Science 252: 668–674

  • Lee SJ, Lee SD, Park JG, Kim CM, Ryu SH, Suh PG . 1995 J. Biol. Chem. 270: 16378–16384

  • Lozano F, Rada C, Jarvis JM, Milstein C . 1993 Nature 363: 271–273

  • Lozano F, Maertzdorf B, Pannell R, Milstein C . 1994 EMBO J. 13: 4617–4622

  • McCafferty J, Hoogenboom HR, Chiswell DJ . 1996 Antibody Engineering: A practical approach IRL Press: London

  • Margolis B, Rhee SG, Felder S, Mervic M, Lyall R, Levitzki A, Ullrich A, Zilberstein A, Schlessinger J . 1989 Cell 57: 1101–1107

  • Meisenhelder J, Suh PG, Rhee SG, Hunter T . 1989 Cell 57: 1109–1122

  • Nebigil CG . 1997 Biochemistry 36: 15949–15958

  • Nieba L, Honegger A, Krebber C, Pluckthun A . 1997 Protein Engng. 10: 435–444

  • Noh DY, Lee YH, Kim SS, Kim YI, Ryu SH, Suh PG, Park JG . 1994 Cancer 73: 36–41

  • Park JG, Lee YH, Kim SS, Park KJ, Noh DY, Ryu SH, Suh PG . 1994 Cancer Res. 54: 2240–2244

  • Rhee SG, Choi KD . 1992 J. Biol. Chem. 267: 12393–12396

  • Rhee SG, Bae YS . 1997 J. Biol. Chem. 272: 15045–15048

  • Richardson JH, Marasco WA . 1995 Trends Biotechnol. 13: 306–310

  • Smith MR, Liu YL, Kim H, Rhee SG, Kung HF . 1990 Science 247: 1074–1077

  • Smith MR, Liu YL, Matthews NT, Rhee SG, Sung WK, Kung HF . 1994 Proc. Natl. Acad. Sci. USA 91: 6554–6558

  • Smith MR, Liu YL, Kim SR, Bae YS, Kim CG, Kwon KS, Rhee SG, Kung HF . 1996 Biochem. Biophys. Res. Commun. 222: 186–193

  • Song C, Hu CD, Masago M, Kariya KI, Yamawaki-Kataoka Y, Shibatohge M, Wu D, Satoh T, Kataoka T . 2001 J. Biol. Chem. 276: 2752–2757

  • Suh PG, Ryu SH, Choi WC, Lee KY, Rhee SG . 1988 J. Biol. Chem. 263: 14497–14504

  • Tsumoto K, Shinjoki K, Kondo H, Uchikawa M, Juji T, Kumagai I . 1998 J. Immunol. Methods 219: 119–129

  • Wetzel R . 1994 Trends Biotechnol. 12: 193–198

  • Yi KS, Chung JH, Kim HJ, Kim IJ, Jung HG, Kim JR, Choi IH, Suh PG, Chung HK . 1999 Hybridoma 18: 243–249

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Acknowledgements

This work was supported by financial support from the Korean Ministry of Education made in the program years of 1997, 21C Frontier Functional Human Genome Project from the Ministry of Science & Technology of Korea, and Critical Technology 21 on ‘Life Phenomena and Function Research’ from the Ministry of Science & Technology of Korea. Lucio Cocco is supported by AIRC, CNR PFBiotec. and FST of Bologna University.

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Authors and Affiliations

  1. Department of Life Science, Division of Molecular and Life Science, Pohang University of Science and Technology, Pohang, Kyungbuk, 790-784, South Korea

    Kye Sook Yi, Hyang Gi Chung, Byung Chang Suh, Euikyung Kim, Sung Ho Ryu & Pann-Ghill Suh

  2. Department of Biochemistry, Seoul National University, College of Medicine, Seoul, 110-799, South Korea

    Jun Ho Chung

  3. Department of Biochemistry, Yeungnam University College of Medicine, 712-749, Taegu, South Korea

    Young Han Lee

  4. Department of Microbiology, Dongguk University College of Medicine, Kyungju, 780-140, South Korea

    Ik Jung Kim

  5. Department of Anatomical Sciences, Cellular Signaling Laboratory, University of Bologna, Via Irnerio, 48 I-40126 Bologna, Italy

    Lucio Cocco

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  1. Kye Sook Yi
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Correspondence to Byung Chang Suh.

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Yi, K., Chung, J., Lee, Y. et al. Inhibition of the EGF-induced activation of phospholipase C-γ1 by a single chain antibody fragment. Oncogene 20, 7954–7964 (2001). https://doi.org/10.1038/sj.onc.1204959

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