Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies

Sweta Iyer, Khatira Anwari, Amber E. Alsop, Wai Shan Yuen, David C.S. Huang, John Carroll, Nicholas A. Smith, Brian J. Smith, Grant Dewson, Ruth M. Kluck

Research output: Contribution to journalArticleResearchpeer-review

Abstract

During apoptosis, Bak and Bax are activated by BH3-only proteins binding to the α2–α5 hydrophobic groove; Bax is also activated via a rear pocket. Here we report that antibodies can directly activate Bak and mitochondrial Bax by binding to the α1–α2 loop. A monoclonal antibody (clone 7D10) binds close to α1 in non-activated Bak to induce conformational change, oligomerization, and cytochrome c release. Anti-FLAG antibodies also activate Bak containing a FLAG epitope close to α1. An antibody (clone 3C10) to the Bax α1–α2 loop activates mitochondrial Bax, but blocks translocation of cytosolic Bax. Tethers within Bak show that 7D10 binding directly extricates α1; a structural model of the 7D10 Fab bound to Bak reveals the formation of a cavity under α1. Our identification of the α1–α2 loop as an activation site in Bak paves the way to develop intrabodies or small molecules that directly and selectively regulate these proteins.
Original languageEnglish
Article number11734
Number of pages10
JournalNature Communications
Volume7
DOIs
Publication statusPublished - 24 May 2016

Keywords

  • apoptosis
  • cancer therapy
  • structural biology

Cite this

Iyer, Sweta ; Anwari, Khatira ; Alsop, Amber E. ; Yuen, Wai Shan ; Huang, David C.S. ; Carroll, John ; Smith, Nicholas A. ; Smith, Brian J. ; Dewson, Grant ; Kluck, Ruth M. / Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies. In: Nature Communications. 2016 ; Vol. 7.
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title = "Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies",
abstract = "During apoptosis, Bak and Bax are activated by BH3-only proteins binding to the α2–α5 hydrophobic groove; Bax is also activated via a rear pocket. Here we report that antibodies can directly activate Bak and mitochondrial Bax by binding to the α1–α2 loop. A monoclonal antibody (clone 7D10) binds close to α1 in non-activated Bak to induce conformational change, oligomerization, and cytochrome c release. Anti-FLAG antibodies also activate Bak containing a FLAG epitope close to α1. An antibody (clone 3C10) to the Bax α1–α2 loop activates mitochondrial Bax, but blocks translocation of cytosolic Bax. Tethers within Bak show that 7D10 binding directly extricates α1; a structural model of the 7D10 Fab bound to Bak reveals the formation of a cavity under α1. Our identification of the α1–α2 loop as an activation site in Bak paves the way to develop intrabodies or small molecules that directly and selectively regulate these proteins.",
keywords = "apoptosis, cancer therapy, structural biology",
author = "Sweta Iyer and Khatira Anwari and Alsop, {Amber E.} and Yuen, {Wai Shan} and Huang, {David C.S.} and John Carroll and Smith, {Nicholas A.} and Smith, {Brian J.} and Grant Dewson and Kluck, {Ruth M.}",
year = "2016",
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Iyer, S, Anwari, K, Alsop, AE, Yuen, WS, Huang, DCS, Carroll, J, Smith, NA, Smith, BJ, Dewson, G & Kluck, RM 2016, 'Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies' Nature Communications, vol. 7, 11734. https://doi.org/10.1038/ncomms11734

Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies. / Iyer, Sweta; Anwari, Khatira; Alsop, Amber E.; Yuen, Wai Shan; Huang, David C.S.; Carroll, John; Smith, Nicholas A.; Smith, Brian J.; Dewson, Grant; Kluck, Ruth M.

In: Nature Communications, Vol. 7, 11734, 24.05.2016.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Identification of an activation site in Bak and mitochondrial Bax triggered by antibodies

AU - Iyer, Sweta

AU - Anwari, Khatira

AU - Alsop, Amber E.

AU - Yuen, Wai Shan

AU - Huang, David C.S.

AU - Carroll, John

AU - Smith, Nicholas A.

AU - Smith, Brian J.

AU - Dewson, Grant

AU - Kluck, Ruth M.

PY - 2016/5/24

Y1 - 2016/5/24

N2 - During apoptosis, Bak and Bax are activated by BH3-only proteins binding to the α2–α5 hydrophobic groove; Bax is also activated via a rear pocket. Here we report that antibodies can directly activate Bak and mitochondrial Bax by binding to the α1–α2 loop. A monoclonal antibody (clone 7D10) binds close to α1 in non-activated Bak to induce conformational change, oligomerization, and cytochrome c release. Anti-FLAG antibodies also activate Bak containing a FLAG epitope close to α1. An antibody (clone 3C10) to the Bax α1–α2 loop activates mitochondrial Bax, but blocks translocation of cytosolic Bax. Tethers within Bak show that 7D10 binding directly extricates α1; a structural model of the 7D10 Fab bound to Bak reveals the formation of a cavity under α1. Our identification of the α1–α2 loop as an activation site in Bak paves the way to develop intrabodies or small molecules that directly and selectively regulate these proteins.

AB - During apoptosis, Bak and Bax are activated by BH3-only proteins binding to the α2–α5 hydrophobic groove; Bax is also activated via a rear pocket. Here we report that antibodies can directly activate Bak and mitochondrial Bax by binding to the α1–α2 loop. A monoclonal antibody (clone 7D10) binds close to α1 in non-activated Bak to induce conformational change, oligomerization, and cytochrome c release. Anti-FLAG antibodies also activate Bak containing a FLAG epitope close to α1. An antibody (clone 3C10) to the Bax α1–α2 loop activates mitochondrial Bax, but blocks translocation of cytosolic Bax. Tethers within Bak show that 7D10 binding directly extricates α1; a structural model of the 7D10 Fab bound to Bak reveals the formation of a cavity under α1. Our identification of the α1–α2 loop as an activation site in Bak paves the way to develop intrabodies or small molecules that directly and selectively regulate these proteins.

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