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

Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing

K.K. Khoo, I. Galleano, F. Gasparri, R. Wieneke, H. Harms, M.H. Poulsen, View ORCID ProfileH.C. Chua, M. Wulf, R. Tampé, View ORCID ProfileS.A. Pless
doi: https://doi.org/10.1101/2020.01.31.925495
K.K. Khoo
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
I. Galleano
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
F. Gasparri
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R. Wieneke
2Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
H. Harms
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
M.H. Poulsen
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
H.C. Chua
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for H.C. Chua
M. Wulf
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R. Tampé
2Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
S.A. Pless
1Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for S.A. Pless
  • For correspondence: stephan.pless@sund.ku.dk
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

Abstract

Manipulation of proteins by chemical modification is a powerful way to decipher their function or harness that function for therapeutic purposes. Despite recent progress in ribosome-dependent and semi-synthetic chemical modifications, these techniques sometimes have limitations in the number and type of modifications that can be simultaneously introduced or their application in live eukaryotic cells. Here we present a new approach to incorporate single or multiple post-translational modifications or non-canonical amino acids into soluble and membrane proteins expressed in eukaryotic cells. We insert synthetic peptides into proteins of interest via tandem protein trans-splicing using two orthogonal split intein pairs and validate our approach by investigating different aspects of GFP, NaV1.5 and P2X2 receptor function. Because the approach can introduce virtually any chemical modification into both intracellular and extracellular regions of target proteins, we anticipate that it will overcome some of the drawbacks of other semi-synthetic or ribosome-dependent methods to engineer proteins.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted February 02, 2020.
Download PDF
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.
Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing
(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
Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing
K.K. Khoo, I. Galleano, F. Gasparri, R. Wieneke, H. Harms, M.H. Poulsen, H.C. Chua, M. Wulf, R. Tampé, S.A. Pless
bioRxiv 2020.01.31.925495; doi: https://doi.org/10.1101/2020.01.31.925495
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing
K.K. Khoo, I. Galleano, F. Gasparri, R. Wieneke, H. Harms, M.H. Poulsen, H.C. Chua, M. Wulf, R. Tampé, S.A. Pless
bioRxiv 2020.01.31.925495; doi: https://doi.org/10.1101/2020.01.31.925495

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

  • Synthetic Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4657)
  • Biochemistry (10309)
  • Bioengineering (7629)
  • Bioinformatics (26217)
  • Biophysics (13462)
  • Cancer Biology (10637)
  • Cell Biology (15354)
  • Clinical Trials (138)
  • Developmental Biology (8461)
  • Ecology (12766)
  • Epidemiology (2067)
  • Evolutionary Biology (16781)
  • Genetics (11368)
  • Genomics (15416)
  • Immunology (10562)
  • Microbiology (25064)
  • Molecular Biology (10165)
  • Neuroscience (54203)
  • Paleontology (398)
  • Pathology (1658)
  • Pharmacology and Toxicology (2878)
  • Physiology (4319)
  • Plant Biology (9206)
  • Scientific Communication and Education (1582)
  • Synthetic Biology (2543)
  • Systems Biology (6759)
  • Zoology (1454)