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Related Experiment Videos

Heparin-aggregated RANTES can be crystallised.

Enrico A Stura1, Loïc Martin, Hugues Lortat-Jacob

  • 1CEA, Départment d'Ingénierie des Protéines (DIEP), CE Saclay, 91191 Gif-sur-Yvette, France. estura@cea.fr

Acta Crystallographica. Section D, Biological Crystallography
|September 28, 2002
PubMed
Summary
This summary is machine-generated.

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Heparin-derived oligosaccharides promote severe aggregation of RANTES (regulated on activation normal T-cell expressed), surprisingly enhancing protein crystallization. Optimal crystal growth was achieved with a six-moiety sugar, challenging prior assumptions about protein aggregation and crystallization suitability.

Area of Science:

  • Structural Biology
  • Protein Crystallography
  • Biochemistry

Background:

  • RANTES (regulated on activation normal T-cell expressed) is a chemokine involved in inflammatory responses.
  • Protein crystallization is crucial for determining protein structure but often challenging for proteins prone to aggregation.
  • Heparin-derived sulphated oligosaccharides are known to interact with chemokines.

Purpose of the Study:

  • To investigate the effect of heparin-derived sulphated oligosaccharides on RANTES crystallization.
  • To determine the optimal conditions for co-crystallizing RANTES with these oligosaccharides.
  • To elucidate the binding interactions between RANTES and sulphated oligosaccharides during crystallization.

Main Methods:

  • Co-crystallization of RANTES with heparin-derived sulphated oligosaccharides of varying lengths (1-12 sugar moieties).

Related Experiment Videos

  • X-ray diffraction analysis of the obtained crystals to determine resolution and quality.
  • Analysis of electron density maps to identify the presence and location of bound oligosaccharides.

    • Main Results:

    • Severe aggregation of RANTES was induced by sulphated oligosaccharides, which paradoxically stimulated nucleation and improved crystal quality.
    • The best crystals were obtained using a six-moiety sugar, diffracting to 1.8 Å resolution.
    • No homogeneous binding of sulphated sugars to RANTES was observed in the final crystal structure, suggesting heterogeneous binding or elimination during growth.

    Conclusions:

    • Protein aggregation, induced by specific oligosaccharides, can be beneficial for crystallisation, challenging the notion that polydispersity hinders crystal formation.
    • The length and stoichiometry of oligosaccharides significantly influence RANTES aggregation, nucleation, and crystal growth.
    • The absence of bound sugars in the crystal structure indicates complex interactions or dynamic processes during crystallization.