Jove
Visualize
Contact Us

Related Experiment Videos

Efficient introduction of alkene functionality into proteins in vivo

J C van Hest1, D A Tirrell

  • 1Department of Polymer Science and Engineering, University of Massachusetts, Amherst 01003, USA.

FEBS Letters
|June 30, 1998
PubMed
Summary

The methionine analogue homoallylglycine (Hag) can replace methionine in Escherichia coli protein synthesis. This enables novel protein modification strategies using the intact Hag vinyl group in purified proteins.

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

Protein-based materials, toward a new level of structural control.

Chemical communications (Cambridge, England)·2002
Same author

Confinement-induced order of tethered alkyl chains at the water/vapor interface.

Physical review. E, Statistical, nonlinear, and soft matter physics·2002
Same author

Engineering the extracellular matrix: a novel approach to polymeric biomaterials. I. Control of the physical properties of artificial protein matrices designed to support adhesion of vascular endothelial cells.

Biomacromolecules·2001
Same author

Thermal and structural properties of biologically derived monodisperse hairy-rod polymers.

Biomacromolecules·2001
Same author

Effects of temperature and pressure on the aggregation properties of an engineered elastin model polypeptide in aqueous solution.

Biomacromolecules·2001
Same author

Biosynthesis of a highly stable coiled-coil protein containing hexafluoroleucine in an engineered bacterial host.

Journal of the American Chemical Society·2001
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

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Synthetic Biology

Background:

  • Methionine is an essential amino acid crucial for protein biosynthesis.
  • Escherichia coli is a widely used host for recombinant protein production.
  • Limited methods exist for post-translational modification of proteins in vivo.

Purpose of the Study:

  • To investigate the utilization of 2-amino-5-hexenoic acid (homoallylglycine, Hag) by Escherichia coli for protein synthesis.
  • To assess the extent of methionine replacement by Hag in a model protein.
  • To explore the potential of Hag for novel protein modification strategies.

Main Methods:

  • Utilizing a methionine auxotroph of Escherichia coli.
  • Supplementing the growth medium with Hag.

Related Experiment Videos

  • Expressing mouse dihydrofolate reductase (mDHFR) under a bacteriophage T5 promoter.
  • Performing N-terminal sequencing to determine initiator site occupancy.
  • Analyzing purified mDHFR for Hag incorporation.

    • Main Results:

    • Escherichia coli successfully incorporated Hag during both initiation and elongation of protein synthesis.
    • Approximately 85% of methionine residues in mDHFR were replaced by Hag.
    • N-terminal sequencing revealed 92+/-5% occupancy of the initiator site by Hag.
    • The vinyl group of Hag remained intact in the purified mDHFR protein.

    Conclusions:

    • Homoallylglycine (Hag) can be efficiently incorporated into proteins by Escherichia coli, replacing native methionine.
    • This Hag incorporation results in proteins with a functional vinyl group, suitable for further chemical modification.
    • The findings open new avenues for bio-orthogonal chemistry and the engineering of novel protein functionalities in bacterial expression systems.