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Telluromethionine in structural biochemistry

J O Boles1, L Lebioda, R B Dunlap

  • 1Department of Chemistry, Tennessee Technological University, Cookeville 38505, USA.

SAAS Bulletin, Biochemistry and Biotechnology
|January 1, 1995
PubMed
Summary
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Researchers partially incorporated tellurium-containing amino acids (telluromethionine) into E. coli dihydrofolate reductase (DHFR). This method aids macromolecular crystallography phasing and provides a probe for nuclear magnetic resonance (NMR) studies.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Biophysics

Background:

  • Solving the phase problem in macromolecular crystallography requires suitable heavy-atom derivatives.
  • Internal labeling of proteins offers a valuable tool for crystallographic phasing and spectroscopic studies.

Purpose of the Study:

  • To demonstrate the partial incorporation of telluromethionine into E. coli dihydrofolate reductase (DHFR) via protein biosynthesis.
  • To evaluate the impact of telluromethionine incorporation on DHFR activity and substrate binding.
  • To establish telluromethionine as a useful tool for both X-ray crystallography and NMR spectroscopy.

Main Methods:

  • Utilizing protein biosynthesis for site-specific incorporation of telluromethionine into E. coli dihydrofolate reductase.
  • Assessing enzyme activity through specific activity measurements.

Related Experiment Videos

  • Determining substrate binding affinity using binding ratio analysis with methotrexate.

    • Main Results:

    • Partial incorporation of telluromethionine into E. coli DHFR was achieved without apparent perturbations to enzyme activity or substrate binding.
    • Enzymes containing two moles of telluromethionine exhibited a specific activity of 42 units/mg.
    • A 1:1 binding ratio with methotrexate was observed for the telluromethionine-containing DHFR.

    Conclusions:

    • Telluromethionine incorporation via protein biosynthesis is a viable method for generating heavy-atom derivatives for crystallography.
    • Telluromethionine serves as an effective internal probe for nuclear magnetic resonance (NMR) studies, enabling structure/function investigations.
    • This approach provides a convenient phasing vehicle for X-ray crystallography and a tool for studying protein structure-function relationships.