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Area of Science:

  • Bioconjugate chemistry
  • Nanoparticle science
  • Structural biology

Background:

  • Protein-spherical nucleic acid conjugates (Pro-SNAs) combine protein cores with oligonucleotide shells, offering potential for cellular applications.
  • The ionic environment significantly impacts protein stability and activity, yet is poorly understood for complex bioconjugates like Pro-SNAs.
  • Existing protein structure determination methods cannot resolve the non-uniform ionic distributions around Pro-SNAs.

Purpose of the Study:

  • To determine the counterion radial distribution profile around Pro-SNAs with high resolution.
  • To investigate the influence of salt concentration on the ionic environment of Pro-SNAs.
  • To establish a method for analyzing the structure and function of Pro-SNAs in various ionic conditions.

Main Methods:

  • In situ anomalous small-angle X-ray scattering (ASAXS) to analyze Pro-SNAs in RbCl solutions.
  • Classical density functional theory (DFT) calculations to model counterion distributions.
  • Small-angle X-ray scattering (SAXS) to determine the radial extension of DNA and linkers.

Main Results:

  • ASAXS and DFT revealed the counterion radial distribution profile around Pro-SNAs at 1 nm resolution.
  • Rb+ cations compensated approximately 90% of the negative charge from DNA and linkers at 50 mM RbCl.
  • DFT predicted overcompensation of DNA charge by Rb+ at salt concentrations above 75 mM.

Conclusions:

  • Solution X-ray scattering combined with DFT can resolve counterion distributions and submolecular features of complex nanoparticles like Pro-SNAs.
  • This approach provides insights into Pro-SNA structure and function in different ionic environments.
  • The findings enable predictions of ionic cloud densities based on salt concentration, DNA grafting density, and length.