title: Multimodal interaction with BCL-2 family proteins underlies the proapoptotic activity of PUMA BH3.
abstract: PUMA is a proapoptotic BCL-2 family member that drives the apoptotic response to a diversity of cellular insults. Deciphering the spectrum of PUMA interactions that confer its context-dependent proapoptotic properties remains a high priority goal. Here, we report the synthesis of PUMA SAHBs, structurally stabilized PUMA BH3 helices that, in addition to broadly targeting antiapoptotic proteins, directly bind to proapoptotic BAX. NMR, photocrosslinking, and biochemical analyses revealed that PUMA SAHBs engage an α1/α6 trigger site on BAX to initiate its functional activation. We further demonstrated that a cell-permeable PUMA SAHB analog induces apoptosis in neuroblastoma cells and, like expressed PUMA protein, engages BCL-2, MCL-1, and BAX. Thus, we find that PUMA BH3 is a dual antiapoptotic inhibitor and proapoptotic direct activator, and its mimetics may serve as effective pharmacologic triggers of apoptosis in resistant human cancers.
authors.affiliation: Department of Pediatric Oncology, Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Edwards
authors.forename: Evripidis
authors.initials: E
authors.lastname: Gavathiotis
authors.forename: James L
authors.initials: JL
authors.lastname: LaBelle
authors.forename: Craig R
authors.initials: CR
authors.lastname: Braun
authors.forename: Kwadwo A
authors.initials: KA
authors.lastname: Opoku-Nsiah
authors.forename: Gregory H
authors.initials: GH
authors.lastname: Bird
authors.forename: Loren D
authors.initials: LD
authors.lastname: Walensky
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000737:chemistry* / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D004789:Enzyme Activation; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016062:Porosity; D017434:Protein Structure, Tertiary; D040901:Proteomics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D053148:Caspase 3; D053179:Caspase 7
references.citation: Cell. 2013 Jan 31;152(3):519-31
references.pmid: 23374347
references.citation: Apoptosis. 2008 Jan;13(1):87-95
references.pmid: 17968660
references.citation: Nat Med. 2008 Feb;14(2):144-53
references.pmid: 18223655
references.citation: Blood. 2005 Dec 15;106(13):4131-8
references.pmid: 16118324
references.citation: J Biomol NMR. 1995 Nov;6(3):277-93
references.pmid: 8520220
references.citation: Cell Death Differ. 2005 Oct;12(10):1310-8
references.pmid: 15905879
references.citation: Science. 2010 Dec 3;330(6009):1390-3
references.pmid: 21127253
references.citation: Nat Chem Biol. 2012 Jul;8(7):639-45
references.pmid: 22634637
references.citation: Nature. 2005 Jun 2;435(7042):677-81
references.pmid: 15902208
references.citation: Proc Natl Acad Sci U S A. 2013 Mar 12;110(11):E986-95
references.pmid: 23404709
references.citation: Cell Cycle. 2013 May 1;12(9):1333-4
references.pmid: 23588072
references.citation: Mol Cell. 2001 Mar;7(3):673-82
references.pmid: 11463391
references.citation: J Biol Chem. 2003 Dec 5;278(49):48935-41
references.pmid: 14522999
references.citation: Cancer Cell. 2002 Sep;2(3):183-92
references.pmid: 12242151
references.citation: J Am Chem Soc. 2007 Mar 7;129(9):2456-7
references.pmid: 17284038
references.citation: Anal Chem. 1995 Apr 15;67(8):1426-36
references.pmid: 7741214
references.citation: Nat Med. 2013 Feb;19(2):202-8
references.pmid: 23291630
references.citation: Cell Death Differ. 2009 Apr;16(4):555-63
references.pmid: 19079139
references.citation: ACS Chem Biol. 2013 Mar 15;8(3):506-12
references.pmid: 23214419
references.citation: J Neurosci. 2007 Nov 21;27(47):12989-99
references.pmid: 18032672
references.citation: Methods Mol Biol. 2004;278:313-52
references.pmid: 15318002
references.citation: J Clin Invest. 2012 Jun;122(6):2018-31
references.pmid: 22622039
references.citation: Mol Cell. 2011 Nov 18;44(4):517-31
references.pmid: 22036586
references.citation: Structure. 2008 May;16(5):818-29
references.pmid: 18462686
references.citation: Mol Cell. 2004 Dec 3;16(5):807-18
references.pmid: 15574335
references.citation: Nature. 2008 Oct 23;455(7216):1076-81
references.pmid: 18948948
references.citation: Sci Transl Med. 2012 Aug 22;4(148):148ra117
references.pmid: 22914623
references.citation: Cancer Res. 2012 May 15;72(10):2565-77
references.pmid: 22589275
references.citation: Cell Death Differ. 2012 Apr;19(4):642-9
references.pmid: 22015606
references.citation: Science. 2007 Feb 9;315(5813):856-9
references.pmid: 17289999
references.citation: J Cell Biol. 2011 Jul 11;194(1):39-48
references.pmid: 21727192
references.citation: Cancer Res. 2008 May 1;68(9):3421-8
references.pmid: 18451170
references.citation: J Neurochem. 2006 Mar;96(5):1213-26
references.pmid: 16478523
references.citation: Curr Top Med Chem. 2011;11(3):281-90
references.pmid: 21320058
references.citation: Nature. 2010 Feb 18;463(7283):899-905
references.pmid: 20164920
references.citation: Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11318-23
references.pmid: 11572983
references.citation: Methods Enzymol. 2008;446:387-408
references.pmid: 18603135
references.citation: J Mol Biol. 2008 Jul 25;380(5):958-71
references.pmid: 18589438
references.citation: Nat Chem Biol. 2013 Mar;9(3):163-8
references.pmid: 23340338
references.citation: Oncogene. 2008 Dec;27 Suppl 1:S71-83
references.pmid: 19641508
references.citation: Cell Death Differ. 2011 Oct;18(10):1545-6
references.pmid: 21909118
references.citation: J Exp Med. 2006 Dec 25;203(13):2939-51
references.pmid: 17178918
references.citation: J Mol Biol. 2008 Dec 12;384(2):313-23
references.pmid: 18835564
references.citation: Immunity. 2012 Sep 21;37(3):451-62
references.pmid: 22960223
references.citation: Science. 2004 Sep 3;305(5689):1466-70
references.pmid: 15353804
references.citation: J Cell Biol. 2003 Aug 18;162(4):587-97
references.pmid: 12913114
references.citation: Cell. 2000 Nov 10;103(4):645-54
references.pmid: 11106734
references.citation: Nat Struct Mol Biol. 2013 May;20(5):589-97
references.pmid: 23604079
references.citation: Nat Chem Biol. 2010 Aug;6(8):595-601
references.pmid: 20562877
references.citation: Mol Cell. 2009 Nov 13;36(3):487-99
references.pmid: 19917256
references.citation: Mol Cell. 2010 Nov 12;40(3):481-92
references.pmid: 21070973
references.citation: Mol Cell. 2001 Mar;7(3):683-94
references.pmid: 11463392
references.citation: Science. 2003 Nov 7;302(5647):1036-8
references.pmid: 14500851
references.citation: Curr Protoc Chem Biol. 2011 Sep 1;3(3):99-117
references.pmid: 23801563
references.citation: J Neurosci. 2006 Jul 5;26(27):7257-64
references.pmid: 16822983
references.citation: J Cell Biol. 2007 Apr 23;177(2):277-87
references.pmid: 17452531
references.citation: Mol Cell. 2006 Oct 20;24(2):199-210
references.pmid: 17052454
references.citation: J Biol Chem. 2011 Jan 7;286(1):491-501
references.pmid: 21041309
references.citation: J Cell Biol. 2009 Apr 20;185(2):279-90
references.pmid: 19380879
references.citation: Mol Cell. 2005 Feb 18;17(4):525-35
references.pmid: 15721256
references.citation: Clin Cancer Res. 2006 May 1;12(9):2928-36
references.pmid: 16675590
references.citation: Methods Enzymol. 2008;446:369-86
references.pmid: 18603134
references.citation: Mol Biol Cell. 2009 Jul;20(13):3077-87
references.pmid: 19439449
references.citation: Methods Enzymol. 2007;430:283-331
references.pmid: 17913643
references.citation: Nat Struct Mol Biol. 2013 May;20(5):536-8
references.pmid: 23649360
references.citation: Chem Biol. 2010 Dec 22;17(12):1325-33
references.pmid: 21168768
references.citation: Cancer Cell. 2003 Oct;4(4):321-8
references.pmid: 14585359
references.citation: Mol Cell. 2006 Dec 8;24(5):677-688
references.pmid: 17157251
references.citation: Cell Death Differ. 2007 Jan;14(1):128-36
references.pmid: 16645638