Dynamic reconfiguration of pro-apoptotic BAK on membranes.
Animals
Binding Sites
Cloning, Molecular
Deuterium Exchange Measurement
Escherichia coli
/ genetics
Gene Expression
Genetic Vectors
/ chemistry
Humans
Kinetics
Liposomes
/ chemistry
Membrane Lipids
/ chemistry
Mice
Models, Molecular
Nuclear Magnetic Resonance, Biomolecular
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Folding
Protein Interaction Domains and Motifs
Protein Multimerization
Proto-Oncogene Proteins c-bcl-2
/ chemistry
Recombinant Proteins
/ chemistry
Thermodynamics
bcl-2 Homologous Antagonist-Killer Protein
/ chemistry
BAK
BCL-2
apoptosis
hydrogen-deuterium exchange mass spectrometry
membrane
Journal
The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664
Informations de publication
Date de publication:
18 10 2021
18 10 2021
Historique:
revised:
02
08
2021
received:
11
11
2020
accepted:
06
08
2021
pubmed:
16
9
2021
medline:
15
12
2021
entrez:
15
9
2021
Statut:
ppublish
Résumé
BAK and BAX, the effectors of intrinsic apoptosis, each undergo major reconfiguration to an activated conformer that self-associates to damage mitochondria and cause cell death. However, the dynamic structural mechanisms of this reconfiguration in the presence of a membrane have yet to be fully elucidated. To explore the metamorphosis of membrane-bound BAK, we employed hydrogen-deuterium exchange mass spectrometry (HDX-MS). The HDX-MS profile of BAK on liposomes comprising mitochondrial lipids was consistent with known solution structures of inactive BAK. Following activation, HDX-MS resolved major reconfigurations in BAK. Mutagenesis guided by our HDX-MS profiling revealed that the BCL-2 homology (BH) 4 domain maintains the inactive conformation of BAK, and disrupting this domain is sufficient for constitutive BAK activation. Moreover, the entire N-terminal region preceding the BAK oligomerisation domains became disordered post-activation and remained disordered in the activated oligomer. Removal of the disordered N-terminus did not impair, but rather slightly potentiated, BAK-mediated membrane permeabilisation of liposomes and mitochondria. Together, our HDX-MS analyses reveal new insights into the dynamic nature of BAK activation on a membrane, which may provide new opportunities for therapeutic targeting.
Identifiants
pubmed: 34523147
doi: 10.15252/embj.2020107237
pmc: PMC8521275
doi:
Substances chimiques
BAK1 protein, human
0
BCL2 protein, human
0
Liposomes
0
Membrane Lipids
0
Proto-Oncogene Proteins c-bcl-2
0
Recombinant Proteins
0
bcl-2 Homologous Antagonist-Killer Protein
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e107237Commentaires et corrections
Type : CommentIn
Informations de copyright
© 2021 The Authors.
Références
Cell Death Differ. 2015 Oct;22(10):1665-75
pubmed: 25744027
Nat Commun. 2015 Jul 24;6:7875
pubmed: 26204806
EMBO J. 2019 Jan 15;38(2):
pubmed: 30573668
Cancer Cell. 2018 Dec 10;34(6):879-891
pubmed: 30537511
Cell Death Differ. 2012 Apr;19(4):661-70
pubmed: 22015607
Nat Commun. 2016 May 24;7:11734
pubmed: 27217060
EMBO J. 2016 Feb 15;35(4):389-401
pubmed: 26783362
Mol Cell. 2021 May 20;81(10):2123-2134.e5
pubmed: 33794146
Nucleic Acids Res. 2020 Jan 8;48(D1):D1145-D1152
pubmed: 31686107
Nat Struct Mol Biol. 2020 Nov;27(11):1024-1031
pubmed: 32929280
Nat Chem Biol. 2019 Apr;15(4):322-330
pubmed: 30718816
Mol Cell. 2006 Dec 8;24(5):677-688
pubmed: 17157251
J Biol Chem. 2010 Jun 4;285(23):17614-27
pubmed: 20382739
Cell. 2013 Jan 31;152(3):519-31
pubmed: 23374347
Nat Chem Biol. 2012 Jul;8(7):639-45
pubmed: 22634637
J Am Soc Mass Spectrom. 2012 Sep;23(9):1512-21
pubmed: 22692830
Nature. 2005 Jun 2;435(7042):677-81
pubmed: 15902208
Nat Chem Biol. 2019 Nov;15(11):1057-1066
pubmed: 31591564
Cell Death Differ. 2014 Feb;21(2):196-205
pubmed: 24162660
Nature. 2016 Oct 27;538(7626):477-482
pubmed: 27760111
EMBO J. 2016 Feb 15;35(4):402-13
pubmed: 26783364
Genes Dev. 2007 Aug 1;21(15):1937-48
pubmed: 17671092
Proc Natl Acad Sci U S A. 2008 Nov 11;105(45):17379-83
pubmed: 18987313
EMBO J. 2005 Jun 15;24(12):2096-103
pubmed: 15920484
Science. 2001 Apr 27;292(5517):727-30
pubmed: 11326099
Nat Rev Mol Cell Biol. 2014 Jan;15(1):49-63
pubmed: 24355989
Mol Cell. 2009 Nov 25;36(4):696-703
pubmed: 19941828
J Biol Chem. 2010 Feb 26;285(9):6636-47
pubmed: 20008353
EMBO J. 2021 Oct 18;40(20):e107237
pubmed: 34523147
J Biol Chem. 2013 Sep 6;288(36):26027-26038
pubmed: 23893415
Genes Dev. 2016 May 15;30(10):1240-50
pubmed: 27198225
J Biol Chem. 1998 Apr 24;273(17):10777-83
pubmed: 9553144
Nat Struct Mol Biol. 2005 Aug;12(8):671-7
pubmed: 16041387
Cell Death Differ. 2008 Oct;15(10):1564-71
pubmed: 18551131
Nat Commun. 2015 Apr 16;6:6841
pubmed: 25880232
Proc Natl Acad Sci U S A. 1999 May 11;96(10):5492-7
pubmed: 10318911
J Cell Biol. 1999 Mar 8;144(5):903-14
pubmed: 10085290
Proteomics. 2019 Mar;19(6):e1800098
pubmed: 30592560
J Biol Chem. 2010 Sep 10;285(37):28924-37
pubmed: 20605789
Oncogene. 2001 Nov 15;20(52):7668-76
pubmed: 11753644
Nat Commun. 2015 Aug 14;6:8042
pubmed: 26271728
Cell Chem Biol. 2017 Apr 20;24(4):493-506.e5
pubmed: 28392146
J Biol Chem. 2004 Jul 16;279(29):30081-91
pubmed: 15138279
Structure. 2018 Oct 2;26(10):1346-1359.e5
pubmed: 30122452
J Biol Chem. 2002 Dec 20;277(51):49360-5
pubmed: 12381734
Mass Spectrom Rev. 2006 Jan-Feb;25(1):158-70
pubmed: 16208684
Mol Cell. 2008 May 9;30(3):369-80
pubmed: 18471982
Science. 2003 Jul 25;301(5632):513-7
pubmed: 12881569
Proc Natl Acad Sci U S A. 2017 Jul 18;114(29):7629-7634
pubmed: 28673969
Nucleic Acids Res. 2019 Jan 8;47(D1):D442-D450
pubmed: 30395289
Mol Cell. 2014 Nov 20;56(4):496-505
pubmed: 25458844
Sci Adv. 2018 Oct 24;4(10):eaau4130
pubmed: 30397651
J Cell Biol. 2007 Apr 23;177(2):277-87
pubmed: 17452531
Elife. 2017 Feb 06;6:
pubmed: 28182867
EMBO J. 2016 Jan 18;35(2):208-36
pubmed: 26702098
Mol Cell. 2014 Sep 18;55(6):938-946
pubmed: 25175025