Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1
Abstract
Overexpression of anti-apoptotic Bcl-2 family proteins contributes to cancer progression and confers resistance to chemotherapy. Small molecules that target Bcl-2 are used in the clinic to treat leukemia, but tight and selective inhibitors are not available for Bcl-2 paralog Bfl-1. Guided by computational analysis, we designed variants of the native BH3 motif PUMA that are > 150-fold selective for Bfl-1 binding. The designed peptides potently trigger disruption of the mitochondrial outer membrane in cells dependent on Bfl-1, but not in cells dependent on other anti-apoptotic homologs. High-resolution crystal structures show that designed peptide FS2 binds Bfl-1 in a shifted geometry, relative to PUMA and other binding partners, due to a set of epistatic mutations. FS2 modified with an electrophile reacts with a cysteine near the peptide-binding groove to augment specificity. Designed Bfl-1 binders provide reagents for cellular profiling and leads for developing enhanced and cell-permeable peptide or small-molecule inhibitors.
- Authors:
-
[1];
[2];
[1]
- Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
- Dana-Farber Cancer Institute, Boston, MA (United States)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of General Medical Sciences; National Inst. of Health; NIH-ORIP HEI; National Science Foundation (NSF); MIT School of Science Ludwig Fund for Cancer Research; Koch Institute/MIT - Dana-Farber/Harvard Cancer Center Bridge Project
- OSTI Identifier:
- 1368344
- Grant/Contract Number:
- AC02-06CH11357; P41 GM103403; S10 RR029205; R01-GM110048
- Resource Type:
- Accepted Manuscript
- Journal Name:
- eLife
- Additional Journal Information:
- Journal Volume: 6; Journal Issue: 2017; Journal ID: ISSN 2050-084X
- Publisher:
- eLife Sciences Publications, Ltd.
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Jenson, Justin M., Ryan, Jeremy A., Grant, Robert A., Letai, Anthony, and Keating, Amy E. Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1. United States: N. p., 2017.
Web. doi:10.7554/eLife.25541.
Jenson, Justin M., Ryan, Jeremy A., Grant, Robert A., Letai, Anthony, & Keating, Amy E. Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1. United States. https://doi.org/10.7554/eLife.25541
Jenson, Justin M., Ryan, Jeremy A., Grant, Robert A., Letai, Anthony, and Keating, Amy E. Thu .
"Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1". United States. https://doi.org/10.7554/eLife.25541. https://www.osti.gov/servlets/purl/1368344.
@article{osti_1368344,
title = {Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1},
author = {Jenson, Justin M. and Ryan, Jeremy A. and Grant, Robert A. and Letai, Anthony and Keating, Amy E.},
abstractNote = {Overexpression of anti-apoptotic Bcl-2 family proteins contributes to cancer progression and confers resistance to chemotherapy. Small molecules that target Bcl-2 are used in the clinic to treat leukemia, but tight and selective inhibitors are not available for Bcl-2 paralog Bfl-1. Guided by computational analysis, we designed variants of the native BH3 motif PUMA that are > 150-fold selective for Bfl-1 binding. The designed peptides potently trigger disruption of the mitochondrial outer membrane in cells dependent on Bfl-1, but not in cells dependent on other anti-apoptotic homologs. High-resolution crystal structures show that designed peptide FS2 binds Bfl-1 in a shifted geometry, relative to PUMA and other binding partners, due to a set of epistatic mutations. FS2 modified with an electrophile reacts with a cysteine near the peptide-binding groove to augment specificity. Designed Bfl-1 binders provide reagents for cellular profiling and leads for developing enhanced and cell-permeable peptide or small-molecule inhibitors.},
doi = {10.7554/eLife.25541},
journal = {eLife},
number = 2017,
volume = 6,
place = {United States},
year = {Thu Jun 08 00:00:00 EDT 2017},
month = {Thu Jun 08 00:00:00 EDT 2017}
}
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