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Related Experiment Videos

Phasing with mercury at 1 A wavelength.

C Dumas1, S Duquerroy, J Janin

  • 1Centre de Biochimie Structurale, U 414 INSERM, UMR 9955 CNRS-Université Montpellier, France.

Acta Crystallographica. Section D, Biological Crystallography
|September 1, 1995
PubMed
Summary

This study demonstrates how tunable synchrotron X-ray sources and heavy-atom replacement effectively determine protein structures. Optimized anomalous scattering with mercury derivatives enabled solving complex protein structures, including nucleoside diphosphate kinase and enolase.

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Area of Science:

  • Structural Biology
  • Biophysics
  • Crystallography

Background:

  • Synchrotron X-ray sources offer tunable beams crucial for optimizing anomalous scattering.
  • Heavy-atom replacement is a key technique in solving protein structures via X-ray crystallography.

Purpose of the Study:

  • To demonstrate the utility of synchrotron radiation for optimizing anomalous scattering in heavy-atom replacement.
  • To solve the protein structures of Dictyostelium discoideum nucleoside diphosphate kinase and lobster enolase.

Main Methods:

  • Utilized synchrotron X-ray sources for tunable wavelength selection.
  • Employed heavy-atom replacement, specifically mercury derivatives, for phase determination.
  • Applied single isomorphous replacement with anomalous scattering (SIRAS) phasing.

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  • Generated interpretable electron-density maps using solvent flattening.

    • Main Results:

    • Successfully solved the structures of two proteins: Dictyostelium discoideum nucleoside diphosphate kinase (17 kDa) and lobster enolase (47 kDa).
    • A single mercury derivative was sufficient for both structures.
    • High-quality electron-density maps were obtained, facilitating structure interpretation.

    Conclusions:

    • Optimized anomalous scattering using synchrotron sources and mercury derivatives is highly efficient for protein structure determination.
    • The method is effective even for proteins with significant molecular weight in the asymmetric unit.
    • SIRAS phasing combined with solvent flattening provides a robust approach for solving complex protein structures.