Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism

Lauren A. Barclay, Thomas E. Wales, Thomas P. Garner, Franziska Wachter, Susan Lee, Rachel M. Guerra, Michelle L. Stewart, Craig R. Braun, Gregory H. Bird, Evripidis Gavathiotis, John R. Engen, Loren D. Walensky

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

BCL-2 is a negative regulator of apoptosis implicated in homeostatic and pathologic cell survival. The canonical anti-apoptotic mechanism involves entrapment of activated BAX by a groove on BCL-2, preventing BAX homo-oligomerization and mitochondrial membrane poration. The BCL-2 BH4 domain also confers anti-apoptotic functionality, but the mechanism is unknown. We find that a synthetic α-helical BH4 domain binds to BAX with nanomolar affinity and independently inhibits the conformational activation of BAX. Hydrogen-deuterium exchange mass spectrometry demonstrated that the N-terminal conformational changes in BAX induced by a triggering BIM BH3 helix were suppressed by the BCL-2 BH4 helix. Structural analyses localized the BH4 interaction site to a groove formed by residues of α1, α1-α2 loop, and α2-α3 and α5-α6 hairpins on the BAX surface. These data reveal a previously unappreciated binding site for targeted inhibition of BAX and suggest that the BCL-2 BH4 domain may participate in apoptosis blockade by a noncanonical interaction mechanism.

Original languageEnglish (US)
Pages (from-to)873-886
Number of pages14
JournalMolecular Cell
Volume57
Issue number5
DOIs
StatePublished - Mar 5 2015

Fingerprint

Apoptosis
Deuterium
Mitochondrial Membranes
Hydrogen
Mass Spectrometry
Cell Survival
Binding Sites

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Barclay, L. A., Wales, T. E., Garner, T. P., Wachter, F., Lee, S., Guerra, R. M., ... Walensky, L. D. (2015). Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. Molecular Cell, 57(5), 873-886. https://doi.org/10.1016/j.molcel.2015.01.014

Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. / Barclay, Lauren A.; Wales, Thomas E.; Garner, Thomas P.; Wachter, Franziska; Lee, Susan; Guerra, Rachel M.; Stewart, Michelle L.; Braun, Craig R.; Bird, Gregory H.; Gavathiotis, Evripidis; Engen, John R.; Walensky, Loren D.

In: Molecular Cell, Vol. 57, No. 5, 05.03.2015, p. 873-886.

Research output: Contribution to journalArticle

Barclay, LA, Wales, TE, Garner, TP, Wachter, F, Lee, S, Guerra, RM, Stewart, ML, Braun, CR, Bird, GH, Gavathiotis, E, Engen, JR & Walensky, LD 2015, 'Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism', Molecular Cell, vol. 57, no. 5, pp. 873-886. https://doi.org/10.1016/j.molcel.2015.01.014
Barclay LA, Wales TE, Garner TP, Wachter F, Lee S, Guerra RM et al. Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. Molecular Cell. 2015 Mar 5;57(5):873-886. https://doi.org/10.1016/j.molcel.2015.01.014
Barclay, Lauren A. ; Wales, Thomas E. ; Garner, Thomas P. ; Wachter, Franziska ; Lee, Susan ; Guerra, Rachel M. ; Stewart, Michelle L. ; Braun, Craig R. ; Bird, Gregory H. ; Gavathiotis, Evripidis ; Engen, John R. ; Walensky, Loren D. / Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. In: Molecular Cell. 2015 ; Vol. 57, No. 5. pp. 873-886.
@article{3445d5390ce742f99fb9896f696b0098,
title = "Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism",
abstract = "BCL-2 is a negative regulator of apoptosis implicated in homeostatic and pathologic cell survival. The canonical anti-apoptotic mechanism involves entrapment of activated BAX by a groove on BCL-2, preventing BAX homo-oligomerization and mitochondrial membrane poration. The BCL-2 BH4 domain also confers anti-apoptotic functionality, but the mechanism is unknown. We find that a synthetic α-helical BH4 domain binds to BAX with nanomolar affinity and independently inhibits the conformational activation of BAX. Hydrogen-deuterium exchange mass spectrometry demonstrated that the N-terminal conformational changes in BAX induced by a triggering BIM BH3 helix were suppressed by the BCL-2 BH4 helix. Structural analyses localized the BH4 interaction site to a groove formed by residues of α1, α1-α2 loop, and α2-α3 and α5-α6 hairpins on the BAX surface. These data reveal a previously unappreciated binding site for targeted inhibition of BAX and suggest that the BCL-2 BH4 domain may participate in apoptosis blockade by a noncanonical interaction mechanism.",
author = "Barclay, {Lauren A.} and Wales, {Thomas E.} and Garner, {Thomas P.} and Franziska Wachter and Susan Lee and Guerra, {Rachel M.} and Stewart, {Michelle L.} and Braun, {Craig R.} and Bird, {Gregory H.} and Evripidis Gavathiotis and Engen, {John R.} and Walensky, {Loren D.}",
year = "2015",
month = "3",
day = "5",
doi = "10.1016/j.molcel.2015.01.014",
language = "English (US)",
volume = "57",
pages = "873--886",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "5",

}

TY - JOUR

T1 - Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism

AU - Barclay, Lauren A.

AU - Wales, Thomas E.

AU - Garner, Thomas P.

AU - Wachter, Franziska

AU - Lee, Susan

AU - Guerra, Rachel M.

AU - Stewart, Michelle L.

AU - Braun, Craig R.

AU - Bird, Gregory H.

AU - Gavathiotis, Evripidis

AU - Engen, John R.

AU - Walensky, Loren D.

PY - 2015/3/5

Y1 - 2015/3/5

N2 - BCL-2 is a negative regulator of apoptosis implicated in homeostatic and pathologic cell survival. The canonical anti-apoptotic mechanism involves entrapment of activated BAX by a groove on BCL-2, preventing BAX homo-oligomerization and mitochondrial membrane poration. The BCL-2 BH4 domain also confers anti-apoptotic functionality, but the mechanism is unknown. We find that a synthetic α-helical BH4 domain binds to BAX with nanomolar affinity and independently inhibits the conformational activation of BAX. Hydrogen-deuterium exchange mass spectrometry demonstrated that the N-terminal conformational changes in BAX induced by a triggering BIM BH3 helix were suppressed by the BCL-2 BH4 helix. Structural analyses localized the BH4 interaction site to a groove formed by residues of α1, α1-α2 loop, and α2-α3 and α5-α6 hairpins on the BAX surface. These data reveal a previously unappreciated binding site for targeted inhibition of BAX and suggest that the BCL-2 BH4 domain may participate in apoptosis blockade by a noncanonical interaction mechanism.

AB - BCL-2 is a negative regulator of apoptosis implicated in homeostatic and pathologic cell survival. The canonical anti-apoptotic mechanism involves entrapment of activated BAX by a groove on BCL-2, preventing BAX homo-oligomerization and mitochondrial membrane poration. The BCL-2 BH4 domain also confers anti-apoptotic functionality, but the mechanism is unknown. We find that a synthetic α-helical BH4 domain binds to BAX with nanomolar affinity and independently inhibits the conformational activation of BAX. Hydrogen-deuterium exchange mass spectrometry demonstrated that the N-terminal conformational changes in BAX induced by a triggering BIM BH3 helix were suppressed by the BCL-2 BH4 helix. Structural analyses localized the BH4 interaction site to a groove formed by residues of α1, α1-α2 loop, and α2-α3 and α5-α6 hairpins on the BAX surface. These data reveal a previously unappreciated binding site for targeted inhibition of BAX and suggest that the BCL-2 BH4 domain may participate in apoptosis blockade by a noncanonical interaction mechanism.

UR - http://www.scopus.com/inward/record.url?scp=84924044826&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84924044826&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2015.01.014

DO - 10.1016/j.molcel.2015.01.014

M3 - Article

C2 - 25684204

AN - SCOPUS:84924044826

VL - 57

SP - 873

EP - 886

JO - Molecular Cell

JF - Molecular Cell

SN - 1097-2765

IS - 5

ER -