Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Screening of hydrocarbon-stapled peptides for inhibition of calcium-triggered exocytosis

Ying Lai, Michael J. Tuvim, Jeremy Leitz, John Peters, Richard A. Pfuetzner, Luis Esquivies, Qiangjun Zhou, Barbara Czako, Jason B. Cross, Philip Jones, Burton F. Dickey, View ORCID ProfileAxel T. Brunger
doi: https://doi.org/10.1101/2022.03.21.484632
Ying Lai
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: brunger@stanford.edu ylai@scu.edu.cn bdickey@mdanderson.org
Michael J. Tuvim
3Department of Pulmonary Medicine, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jeremy Leitz
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John Peters
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard A. Pfuetzner
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
2Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Luis Esquivies
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
2Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Qiangjun Zhou
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Barbara Czako
4Institute for Applied Cancer Science, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jason B. Cross
4Institute for Applied Cancer Science, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Philip Jones
4Institute for Applied Cancer Science, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Burton F. Dickey
3Department of Pulmonary Medicine, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: brunger@stanford.edu ylai@scu.edu.cn bdickey@mdanderson.org
Axel T. Brunger
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305, USA
2Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Axel T. Brunger
  • For correspondence: brunger@stanford.edu ylai@scu.edu.cn bdickey@mdanderson.org
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

The so-called primary interface between the SNARE complex and synaptotagmin-1 (Syt1) is essential for Ca2+-triggered neurotransmitter release in neuronal synapses. The interacting residues of the primary interface are conserved across different species for synaptotagmins (Syt1, Syt2, Syt9), SNAP-25, and syntaxin-1A homologs involved in fast synchronous release. This Ca2+-independent interface forms prior to Ca2+-triggering and plays a role in synaptic vesicle priming. This primary interface is also conserved in the fusion machinery that is responsible for mucin granule membrane fusion. Ca2+-stimulated mucin secretion is mediated by the SNAREs syntaxin-3, SNAP-23, VAMP8, synaptotagmin-2, and other proteins. Here, we designed and screened a series of hydrocarbon-stapled peptides consisting of SNAP-25 fragments that included some of the key residues involved in the primary interface as observed in high-resolution crystal structures. We selected a subset of four stapled peptides that were highly α-helical as assessed by circular dichroism and that inhibited both Ca2+-independent and Ca2+-triggered ensemble lipid-mixing with neuronal SNAREs and Syt1. In a single-vesicle contentmixing assay with reconstituted neuronal SNAREs and synaptotagmin-1 or with reconstituted airway SNAREs and synaptotagmin-2, the selected peptides also suppressed Ca2+-triggered fusion. Taken together, hydrocarbon-stapled peptides that interfere with the primary interface consequently inhibit Ca2+-triggered exocytosis. Our inhibitor screen suggests that these compounds may be useful to combat mucus hypersecretion that is a major cause of airway obstruction in the pathophysiology of COPD, asthma and cystic fibrosis.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • additional minor corrections

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Back to top
PreviousNext
Posted May 11, 2022.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Screening of hydrocarbon-stapled peptides for inhibition of calcium-triggered exocytosis
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Screening of hydrocarbon-stapled peptides for inhibition of calcium-triggered exocytosis
Ying Lai, Michael J. Tuvim, Jeremy Leitz, John Peters, Richard A. Pfuetzner, Luis Esquivies, Qiangjun Zhou, Barbara Czako, Jason B. Cross, Philip Jones, Burton F. Dickey, Axel T. Brunger
bioRxiv 2022.03.21.484632; doi: https://doi.org/10.1101/2022.03.21.484632
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Screening of hydrocarbon-stapled peptides for inhibition of calcium-triggered exocytosis
Ying Lai, Michael J. Tuvim, Jeremy Leitz, John Peters, Richard A. Pfuetzner, Luis Esquivies, Qiangjun Zhou, Barbara Czako, Jason B. Cross, Philip Jones, Burton F. Dickey, Axel T. Brunger
bioRxiv 2022.03.21.484632; doi: https://doi.org/10.1101/2022.03.21.484632

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Pharmacology and Toxicology
Subject Areas
All Articles
  • Animal Behavior and Cognition (3514)
  • Biochemistry (7367)
  • Bioengineering (5347)
  • Bioinformatics (20326)
  • Biophysics (10046)
  • Cancer Biology (7777)
  • Cell Biology (11353)
  • Clinical Trials (138)
  • Developmental Biology (6453)
  • Ecology (9980)
  • Epidemiology (2065)
  • Evolutionary Biology (13357)
  • Genetics (9373)
  • Genomics (12614)
  • Immunology (7725)
  • Microbiology (19104)
  • Molecular Biology (7465)
  • Neuroscience (41153)
  • Paleontology (301)
  • Pathology (1235)
  • Pharmacology and Toxicology (2142)
  • Physiology (3180)
  • Plant Biology (6880)
  • Scientific Communication and Education (1276)
  • Synthetic Biology (1900)
  • Systems Biology (5328)
  • Zoology (1091)