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

Kye Sook Yi; Jun Ho Chung; Young Han Lee; Hyang Gi Chung; Ik Jung Kim; Byung Chang Suh; Euikyung Kim; Lucio Cocco; Sung Ho Ryu; Pann Ghill Suh

doi:10.1038/sj.onc.1204959

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

Kye Sook Yi, Jun Ho Chung, Young Han Lee, Hyang Gi Chung, Ik Jung Kim, Byung Chang Suh, Euikyung Kim, Lucio Cocco, Sung Ho Ryu, Pann Ghill Suh

Department of Brain Sciences

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

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 (K_d= 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.

Original language	English
Pages (from-to)	7954-7964
Number of pages	11
Journal	Oncogene
Volume	20
Issue number	55
DOIs	https://doi.org/10.1038/sj.onc.1204959
State	Published - 29 Nov 2001

Bibliographical note

Funding Information:
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.

Keywords

EGF
PLC-γ1
Single chain antibody (scFv)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/sj.onc.1204959

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

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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author = "Yi, {Kye Sook} and Chung, {Jun Ho} and Lee, {Young Han} and Chung, {Hyang Gi} and Kim, {Ik Jung} and Suh, {Byung Chang} and Euikyung Kim and Lucio Cocco and Ryu, {Sung Ho} and Suh, {Pann Ghill}",

note = "Funding Information: 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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AU - Chung, Jun Ho

AU - Lee, Young Han

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AU - Kim, Ik Jung

AU - Suh, Byung Chang

AU - Kim, Euikyung

AU - Cocco, Lucio

AU - Ryu, Sung Ho

AU - Suh, Pann Ghill

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PY - 2001/11/29

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N2 - 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.

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

Abstract

Bibliographical note

Keywords

UN SDGs

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