Cellular Uptake and Ultrastructural Localization Underlie the Pro-apoptotic Activity of a Hydrocarbon-stapled BIM BH3 Peptide

Overview

Journal ACS Chem Biol

Publisher American Chemical Society

Specialties Biochemistry
Biology

Date 2015 Jul 8

PMID 26151238

Citations 21

Authors

Amanda L Edwards

Franziska Wachter

Margaret Lammert

Annissa J Huhn

James Luccarelli

Gregory H Bird

Loren D Walensky

Affiliations

Soon will be listed here.

Abstract

Hydrocarbon stapling has been applied to restore and stabilize the α-helical structure of bioactive peptides for biochemical, structural, cellular, and in vivo studies. The peptide sequence, in addition to the composition and location of the installed staple, can dramatically influence the properties of stapled peptides. As a result, constructs that appear similar can have distinct functions and utilities. Here, we perform a side-by-side comparison of stapled peptides modeled after the pro-apoptotic BIM BH3 helix to highlight these principles. We confirm that replacing a salt-bridge with an i, i + 4 hydrocarbon staple does not impair target binding affinity and instead can yield a biologically and pharmacologically enhanced α-helical peptide ligand. Importantly, we demonstrate by electron microscopy that the pro-apoptotic activity of a stapled BIM BH3 helix correlates with its capacity to achieve cellular uptake without membrane disruption and accumulate at the organellar site of mechanistic activity.

Citing Articles

Exploring the Chemical Features and Biomedical Relevance of Cell-Penetrating Peptides.

Moreno-Vargas L, Prada-Gracia D Int J Mol Sci. 2025; 26(1.

PMID: 39795918 PMC: 11720145. DOI: 10.3390/ijms26010059.

Histidine-Covalent Stapled Alpha-Helical Peptides Targeting hMcl-1.

Alboreggia G, Udompholkul P, Baggio C, Muzzarelli K, Assar Z, Pellecchia M J Med Chem. 2024; 67(10):8172-8185.

PMID: 38695666 PMC: 11129181. DOI: 10.1021/acs.jmedchem.4c00277.

Cyclic peptides discriminate BCL-2 and its clinical mutants from BCL-X by engaging a single-residue discrepancy.

Li F, Liu J, Liu C, Liu Z, Peng X, Huang Y Nat Commun. 2024; 15(1):1476.

PMID: 38368459 PMC: 10874388. DOI: 10.1038/s41467-024-45848-1.

Inhibition of FOXP3 by stapled alpha-helical peptides dampens regulatory T cell function.

Hawley K, Eclov R, Schnorenberg M, Tian Y, Shah R, Thomas-Toth A Proc Natl Acad Sci U S A. 2022; 119(42):e2209044119.

PMID: 36227917 PMC: 9586281. DOI: 10.1073/pnas.2209044119.

Harnessing the Therapeutic Potential of Biomacromolecules through Intracellular Delivery of Nucleic Acids, Peptides, and Proteins.

Tian Y, Tirrell M, LaBelle J Adv Healthc Mater. 2022; 11(12):e2102600.

PMID: 35285167 PMC: 9232950. DOI: 10.1002/adhm.202102600.

References

Li Y, Rodewald L, Hoppmann C, Wong E, Lebreton S, Safar P . A versatile platform to analyze low-affinity and transient protein-protein interactions in living cells in real time. Cell Rep. 2014; 9(5):1946-1958. PMC: 4269221. DOI: 10.1016/j.celrep.2014.10.058. View

Chen L, Willis S, Wei A, Smith B, Fletcher J, Hinds M . Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell. 2005; 17(3):393-403. DOI: 10.1016/j.molcel.2004.12.030. View

Czabotar P, Westphal D, Dewson G, Ma S, Hockings C, Fairlie W . Bax crystal structures reveal how BH3 domains activate Bax and nucleate its oligomerization to induce apoptosis. Cell. 2013; 152(3):519-31. DOI: 10.1016/j.cell.2012.12.031. View

Leshchiner E, Parkhitko A, Bird G, Luccarelli J, Bellairs J, Escudero S . Direct inhibition of oncogenic KRAS by hydrocarbon-stapled SOS1 helices. Proc Natl Acad Sci U S A. 2015; 112(6):1761-6. PMC: 4330742. DOI: 10.1073/pnas.1413185112. View

Leshchiner E, Braun C, Bird G, Walensky L . Direct activation of full-length proapoptotic BAK. Proc Natl Acad Sci U S A. 2013; 110(11):E986-95. PMC: 3600461. DOI: 10.1073/pnas.1214313110. View

Bernal F, Tyler A, Korsmeyer S, Walensky L, Verdine G . Reactivation of the p53 tumor suppressor pathway by a stapled p53 peptide. J Am Chem Soc. 2007; 129(9):2456-7. PMC: 6333086. DOI: 10.1021/ja0693587. View

Guharoy M, Chakrabarti P . Secondary structure based analysis and classification of biological interfaces: identification of binding motifs in protein-protein interactions. Bioinformatics. 2007; 23(15):1909-18. DOI: 10.1093/bioinformatics/btm274. View

Bird G, Irimia A, Ofek G, Kwong P, Wilson I, Walensky L . Stapled HIV-1 peptides recapitulate antigenic structures and engage broadly neutralizing antibodies. Nat Struct Mol Biol. 2014; 21(12):1058-67. PMC: 4304871. DOI: 10.1038/nsmb.2922. View

LaBelle J, Katz S, Bird G, Gavathiotis E, Stewart M, Lawrence C . A stapled BIM peptide overcomes apoptotic resistance in hematologic cancers. J Clin Invest. 2012; 122(6):2018-31. PMC: 3366394. DOI: 10.1172/JCI46231. View

10.

Kim W, Bird G, Neff T, Guo G, Kerenyi M, Walensky L . Targeted disruption of the EZH2-EED complex inhibits EZH2-dependent cancer. Nat Chem Biol. 2013; 9(10):643-50. PMC: 3778130. DOI: 10.1038/nchembio.1331. View

11.

Gavathiotis E, Suzuki M, Davis M, Pitter K, Bird G, Katz S . BAX activation is initiated at a novel interaction site. Nature. 2008; 455(7216):1076-81. PMC: 2597110. DOI: 10.1038/nature07396. View

12.

Takada K, Zhu D, Bird G, Sukhdeo K, Zhao J, Mani M . Targeted disruption of the BCL9/β-catenin complex inhibits oncogenic Wnt signaling. Sci Transl Med. 2012; 4(148):148ra117. PMC: 3631420. DOI: 10.1126/scitranslmed.3003808. View

13.

Szlyk B, Braun C, Ljubicic S, Patton E, Bird G, Osundiji M . A phospho-BAD BH3 helix activates glucokinase by a mechanism distinct from that of allosteric activators. Nat Struct Mol Biol. 2013; 21(1):36-42. PMC: 4084830. DOI: 10.1038/nsmb.2717. View

14.

Okamoto T, Segal D, Zobel K, Fedorova A, Yang H, Fairbrother W . Further insights into the effects of pre-organizing the BimBH3 helix. ACS Chem Biol. 2014; 9(3):838-9. DOI: 10.1021/cb400638p. View

15.

Bird G, Madani N, Perry A, Princiotto A, Supko J, He X . Hydrocarbon double-stapling remedies the proteolytic instability of a lengthy peptide therapeutic. Proc Natl Acad Sci U S A. 2010; 107(32):14093-8. PMC: 2922607. DOI: 10.1073/pnas.1002713107. View

16.

Barclay L, Wales T, Garner T, Wachter F, Lee S, Guerra R . Inhibition of Pro-apoptotic BAX by a noncanonical interaction mechanism. Mol Cell. 2015; 57(5):873-886. PMC: 4384643. DOI: 10.1016/j.molcel.2015.01.014. View

17.

Pitter K, Bernal F, LaBelle J, Walensky L . Dissection of the BCL-2 family signaling network with stabilized alpha-helices of BCL-2 domains. Methods Enzymol. 2008; 446:387-408. PMC: 6317341. DOI: 10.1016/S0076-6879(08)01623-6. View

18.

Lau Y, de Andrade P, Wu Y, Spring D . Peptide stapling techniques based on different macrocyclisation chemistries. Chem Soc Rev. 2014; 44(1):91-102. DOI: 10.1039/c4cs00246f. View

19.

Walensky L, Kung A, Escher I, Malia T, Barbuto S, Wright R . Activation of apoptosis in vivo by a hydrocarbon-stapled BH3 helix. Science. 2004; 305(5689):1466-70. PMC: 1360987. DOI: 10.1126/science.1099191. View

20.

Bird G, Boyapalle S, Wong T, Opoku-Nsiah K, Bedi R, Crannell W . Mucosal delivery of a double-stapled RSV peptide prevents nasopulmonary infection. J Clin Invest. 2014; 124(5):2113-24. PMC: 4001541. DOI: 10.1172/JCI71856. View