Computationally designed high specificity inhibitors delineate the roles of BCL2 family proteins in cancer.
Many cancers overexpress one or more of the six human pro-survival BCL2 family proteins to evade apoptosis. To determine which BCL2 protein or proteins block apoptosis in different cancers, we computationally designed three-helix bundle protein inhibitors specific for each BCL2 pro-survival protein. Following in vitro optimization, each inhibitor binds its target with high picomolar to low nanomolar affinity and at least 300-fold specificity. Expression of the designed inhibitors in human cancer cell lines revealed unique dependencies on BCL2 proteins for survival which could not be inferred from other BCL2 profiling methods. Our results show that designed inhibitors can be generated for each member of a closely-knit protein family to probe the importance of specific protein-protein interactions in complex biological processes.
Department of Bioengineering, University of Washington, Seattle, United States.
Stephanie
0000-0002-3738-5907
S
Berger
Department of Biochemistry, University of Washington, Seattle, United States.|Department of Biochemistry, University of Illinois, Urbana, United States.
Erik
E
Procko
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, United States.|Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
Daciana
D
Margineantu
Department of Chemistry and Physics, LaTrobe Institute for Molecular Science, Melbourne, Australia.|Olivia Newton-John Cancer Research Institute, Olivia Newton-John Cancer and Wellness Centre, Heidelberg, Australia.|School of Cancer Medicine, La Trobe University, Melbourne, Australia.|The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.|Department of Medical Biology, University of Melbourne, Parkville, Australia.
Erinna F
EF
Lee
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
Betty W
BW
Shen
Department of Biochemistry, University of Washington, Seattle, United States.
Alex
A
Zelter
Department of Biochemistry, University of Washington, Seattle, United States.|Institute for Protein Design, University of Washington, Seattle, United States.
Daniel-Adriano
DA
Silva
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, United States.|Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
Kusum
K
Chawla
The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.|Department of Medical Biology, University of Melbourne, Parkville, Australia.
Marco J
MJ
Herold
The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.|Department of Medical Biology, University of Melbourne, Parkville, Australia.
Jean-Marc
JM
Garnier
Department of Genome Sciences, University of Washington, Seattle, United States.
Richard
R
Johnson
Department of Genome Sciences, University of Washington, Seattle, United States.
Michael J
MJ
MacCoss
The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.|Department of Medical Biology, University of Melbourne, Parkville, Australia.|Department of Pharmacology and Therapeutics, University of Melbourne, Parkville, Australia.
Guillaume
0000-0002-1193-8147
G
Lessene
Department of Biochemistry, University of Washington, Seattle, United States.
Trisha N
0000-0003-4797-3152
TN
Davis
Department of Bioengineering, University of Washington, Seattle, United States.
Patrick S
PS
Stayton
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
Barry L
BL
Stoddard
Department of Chemistry and Physics, LaTrobe Institute for Molecular Science, Melbourne, Australia.|Olivia Newton-John Cancer Research Institute, Olivia Newton-John Cancer and Wellness Centre, Heidelberg, Australia.|School of Cancer Medicine, La Trobe University, Melbourne, Australia.|The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.|Department of Medical Biology, University of Melbourne, Parkville, Australia.
W Douglas
WD
Fairlie
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, United States.|Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
David M
DM
Hockenbery
Department of Biochemistry, University of Washington, Seattle, United States.|Institute for Protein Design, University of Washington, Seattle, United States.|Howard Hughes Medical Institute, University of Washington, Seattle, United States.
David
0000-0001-7896-6217
D
Baker
D017209:Apoptosis / Q000235:genetics; D019295:Computational Biology; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D006801:Humans; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology; D017433:Protein Structure, Secondary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000235:genetics*
D019253:Proto-Oncogene Proteins c-bcl-2
Bcl-2; cancer; cancer biology; computational biology; computational protein design; human; systems biology