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

Expanding pyrimidine diphosphosugar libraries via structure-based nucleotidylyltransferase engineering.

William A Barton1, John B Biggins, Jiqing Jiang

  • 1Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

Proceedings of the National Academy of Sciences of the United States of America
|October 11, 2002
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

Tetracenomycin Aglycones Primarily Inhibit Cell Growth and Proliferation in Mammalian Cancer Cell Lines.

Applied sciences (Basel, Switzerland)·2026
Same author

Bridging Tumor Growth Dynamics and Survival in Preclinical Oncology Drug Development - A Translational Tumor Growth Inhibition and Time to Event (TGI-TTE) Modeling Framework.

Research square·2026
Same author

Physiologically Based Pharmacokinetic Modeling to Predict Human Pharmacokinetics of a Novel Mithramycin Analog for Ewing Sarcoma.

Research square·2026
Same author

Correction to: Surgical screening protocol for craniocervical instability secondary to ehlers-danlos syndrome and other connective tissue disorders: analysis of a 347 patient case series.

Neurosurgical review·2026
Same author

Characterization of the D8P1C1 Anti-ADAM17 Inhibitory Monoclonal Antibody and Generation of Its Bispecific T-Cell Engager Derivative.

International journal of molecular sciences·2026
Same author

Nonlinear Mixed-Effects Modeling to Characterize the Pharmacokinetics of a Novel Mithramycin Analogue for Ewing Sarcoma in Mice.

Research square·2026

This study engineered enzymes for glycorandomization, expanding the range of available sugars for creating diverse glycosylated biomolecules. A key mutation enhanced enzyme acceptance of various alpha-d-hexopyranosyl phosphates.

Area of Science:

  • Biochemistry
  • Enzymology
  • Synthetic Biology

Background:

  • In vitro glycorandomization is a chemoenzymatic method for producing diverse glycosylated biomolecules from natural product scaffolds.
  • This technique utilizes engineered enzymes, specifically nucleotidylyltransferases and glycosyltransferases, involved in metabolite glycosylation.

Purpose of the Study:

  • To expand the variety of UDP/dTDP sugars usable in glycorandomization.
  • To engineer Salmonella enterica LT2 alpha-d-glucopyranosyl phosphate thymidylyltransferase (E(p)) to accept a broader range of alpha-d-hexopyranosyl phosphates.

Main Methods:

  • Employed a structure-based engineering approach to modify the enzyme E(p).
  • Investigated the design rationale, substrate specificity, and structural characteristics of engineered mutations.

Related Experiment Videos

  • Utilized X-ray crystallography for structural elucidation of enzyme-substrate interactions.

    • Main Results:

    • Identified three engineered mutations with altered substrate specificities.
    • A single amino acid substitution (L89T) significantly broadened the enzyme's acceptance of alpha-d-hexopyranosyl phosphates, including alpha-d-allo-, alpha-d-altro-, and alpha-d-talopyranosyl phosphate.
    • Demonstrated successful and some unexpected outcomes in altering nucleotidylyltransferase specificity through rational design.

    Conclusions:

    • The study provides a blueprint for rationally altering nucleotidylyltransferase specificity.
    • This engineered enzyme expands the toolkit for in vitro glycorandomization.
    • The findings represent a crucial step towards more versatile chemoenzymatic synthesis of complex carbohydrates.