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 Experiment Videos

Expressed protein ligation: a general method for protein engineering

T W Muir1, D Sondhi, P A Cole

  • 1Laboratory of Synthetic Protein Chemistry, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. muirt@rockvax.rockefeller.edu

Proceedings of the National Academy of Sciences of the United States of America
|June 17, 1998
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Correction to: Three‑dimensional topography of scapular nutrient foramina.

Surgical and radiologic anatomy : SRA·2020
Same author

Three-dimensional topography of scapular nutrient foramina.

Surgical and radiologic anatomy : SRA·2020
Same author

Radioiodinated Capsids Facilitate In Vivo Non-Invasive Tracking of Adeno-Associated Gene Transfer Vectors.

Scientific reports·2017
Same author

Access to site-specific Fc-cRGD peptide conjugates through streamlined expressed protein ligation.

Organic & biomolecular chemistry·2016
Same author

LSD1 Histone Demethylase Assays and Inhibition.

Methods in enzymology·2016
Same author

Brain Region-Specific Degeneration with Disease Progression in Late Infantile Neuronal Ceroid Lipofuscinosis (CLN2 Disease).

AJNR. American journal of neuroradiology·2016
Same journal

Chemotactic self-organization captures the dynamics of mammalian hair follicle patterning.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Tomographic imaging of superconducting order using particle-hole interference.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Inhibitory potential of autologous neutralizing antibodies sets quantitative limits on the rebound-competent HIV-1 reservoir.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Inferring epidemiological parameters under an infectious phylogeography model with visitor dynamics.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Analytical modeling for suction cup designs for skin-interfaced wearable devices.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Improving cell-free metabolism through direct integration of artificial respiratory chains.

Proceedings of the National Academy of Sciences of the United States of America·2026
See all related articles

A novel protein ligation method enables precise peptide attachment to recombinant proteins. This technique successfully modified C-terminal Src kinase (Csk), enhancing its catalytic activity and revealing new protein engineering possibilities.

Area of Science:

  • Biochemistry
  • Protein Engineering
  • Chemical Biology

Background:

  • Protein semisynthesis is crucial for creating modified proteins.
  • Expressed protein ligation (EPL) offers a chemoselective approach for protein modification.
  • Post-translational modifications significantly impact protein function.

Purpose of the Study:

  • To describe and validate a protein semisynthesis method using expressed protein ligation.
  • To investigate the functional consequences of tail-phosphorylating C-terminal Src kinase (Csk).
  • To demonstrate the utility of EPL for introducing modifications into proteins.

Main Methods:

  • Expressed protein ligation (EPL) was employed for chemoselective peptide ligation.
  • A phosphotyrosine peptide was ligated to the C terminus of recombinant Csk via a thioester intermediate.

Related Experiment Videos

  • The functional properties of the semisynthetically modified Csk were analyzed.

    • Main Results:

    • Near quantitative ligation of the phosphotyrosine peptide to Csk was achieved.
    • Semisynthetic tail-phosphorylated Csk exhibited an intramolecular phosphotyrosine-Src homology 2 interaction.
    • Catalytic phosphoryl transfer efficiency of Csk increased significantly upon tail-phosphorylation.

    Conclusions:

    • Expressed protein ligation is a powerful and versatile method for protein engineering.
    • This technique allows the introduction of unnatural amino acids, posttranslational modifications, and probes.
    • Tail-phosphorylation of Csk via EPL enhances its catalytic activity and reveals functional insights.