Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA Splicing01:32

RNA Splicing

58.2K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
58.2K
Alternative RNA Splicing02:18

Alternative RNA Splicing

22.8K
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
22.8K
Alternative RNA Splicing02:18

Alternative RNA Splicing

4.3K
4.3K
Pre-mRNA Processing: RNA Splicing01:36

Pre-mRNA Processing: RNA Splicing

6.0K
6.0K
Cotranslational Protein Translocation01:20

Cotranslational Protein Translocation

8.6K
Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
8.6K
Insertion of Single-pass Transmembrane Proteins in the RER01:26

Insertion of Single-pass Transmembrane Proteins in the RER

13.7K
Integral membrane proteins are proteins adhered to the lipid bilayer of a cell organelle or membrane. They can be of two types: transmembrane integral proteins that span the lipid bilayer and monotopic proteins that are attached to either side of the membrane but do not pass through it.
Integral transmembrane proteins possess transmembrane and extra membrane domains. The transmembrane domains are primarily made of 20-25 hydrophobic amino acids arranged in a helical secondary confirmation. These...
13.7K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Visible-Light-Induced Flavin Catalysis: A Green Route to Naphtho[2,1-<i>b</i>]furans via an <i>o</i>-(Naphtho)quinone Intermediate.

Organic letters·2025
Same author

Conformational plasticity of human acid-sensing ion channel 1a.

bioRxiv : the preprint server for biology·2024
Same author

Tribromide enabled step-up generation of spirolactams from esters employing oxidative dearomatization of arenols.

Chemical communications (Cambridge, England)·2024
Same author

Unplugging lateral fenestrations of NALCN reveals a hidden drug binding site within the pore region.

Proceedings of the National Academy of Sciences of the United States of America·2024
Same author

BF<sub>3</sub>·Et<sub>2</sub>O-assisted synthesis of sulfinylated spiro[5.5]trienones from biaryl ynones.

Organic & biomolecular chemistry·2024
Same author

Unplugging lateral fenestrations of NALCN reveals a hidden drug binding site within the pore module.

bioRxiv : the preprint server for biology·2024

Related Experiment Video

Updated: Nov 2, 2025

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

9.1K

Ion channel engineering using protein trans-splicing.

Debayan Sarkar1, Hendrik Harms1, Iacopo Galleano1

  • 1Department of Drug Design and Pharmacology, University of Copenhagen, Copenhagen, Denmark.

Methods in Enzymology
|June 14, 2021
PubMed
Summary

Semi-synthesis using protein trans-splicing (PTS) enables diverse chemical modifications in ion channels. This method allows incorporating non-canonical amino acids and other labels for enhanced functional studies.

Keywords:
Chemical modificationMembrane proteinsProtein semi-synthesis

More Related Videos

Controllable Ion Channel Expression through Inducible Transient Transfection
10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

9.6K
Using the E1A Minigene Tool to Study mRNA Splicing Changes
10:25

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

5.2K

Related Experiment Videos

Last Updated: Nov 2, 2025

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

9.1K
Controllable Ion Channel Expression through Inducible Transient Transfection
10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

9.6K
Using the E1A Minigene Tool to Study mRNA Splicing Changes
10:25

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

5.2K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biophysics

Background:

  • Site-directed mutagenesis and genetic code expansion are key for membrane protein functional studies.
  • Semi-synthetic strategies allow incorporating a wide array of chemical modifications into proteins.
  • Protein trans-splicing (PTS) is a powerful technique for protein reconstitution in live cells.

Purpose of the Study:

  • To explore the application of protein trans-splicing (PTS) and tandem PTS (tPTS) for semi-synthesis of ion channels.
  • To incorporate non-canonical amino acids (ncAAs) and post-translational modifications into ion channels using semi-synthesis.
  • To expand the repertoire of modifications available for ion channel research.

Main Methods:

  • Utilized protein trans-splicing (PTS) and tandem PTS (tPTS) for semi-synthetic protein reconstitution.
  • Employed Xenopus laevis oocytes as a system for ion channel semi-synthesis.
  • Incorporated non-canonical amino acids (ncAAs) and post-translational modification mimics.

Main Results:

  • Demonstrated the successful semi-synthesis of ion channels using PTS and tPTS in Xenopus laevis oocytes.
  • Successfully incorporated various chemical modifications, including ncAAs and their mimics.
  • Validated the utility of PTS-based semi-synthesis for introducing diverse modifications.

Conclusions:

  • Protein trans-splicing (PTS) offers a versatile platform for the semi-synthesis of ion channels.
  • This strategy significantly broadens the scope of chemical modifications applicable to ion channel research.
  • The developed approach holds potential for deeper functional and pharmacological insights into ion channels.