Dynamic light-scattering analysis of the electrostatic interaction of hexahistidine-tagged cytochrome P450 enzyme with semiconductor quantum dots

Chemphyschem. 2006 May 12;7(5):1112-8. doi: 10.1002/cphc.200500660.

Abstract

Currently, there is great interest in the development of methods suitable for determining the stoichiometry of biomolecules attached to nanoparticles. We describe the use of the dynamic light-scattering technique (DLS) to determine the stoichiometry of the protein cytochrome P450(BSbeta) attached to CdS and CdSe quantum dots (QDs). The enzyme-conjugated QDs have different diffusion characteristics compared to the QD and enzyme precursors, expressed in their size, scattering intensity as well as zeta-potential values. The significant enhancement of the scattering intensity of QDs observed upon conjugation with the P450(BSbeta) due to the refractive-index increment and the systematic variation in zeta potential resulting from charge neutralization of the anionic QDs by the cationic histidine-tagged P450(BSbeta) have been used for stoichiometry determination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anions
  • Biophysics / methods*
  • Cations
  • Chemistry, Physical / methods
  • Cytochrome P-450 Enzyme System / chemistry*
  • Enzymes / chemistry
  • Escherichia coli / metabolism
  • Histidine / chemistry*
  • Light
  • Molecular Conformation
  • Oligopeptides / chemistry*
  • Quantum Dots*
  • Scattering, Radiation
  • Semiconductors*
  • Static Electricity

Substances

  • Anions
  • Cations
  • Enzymes
  • His-His-His-His-His-His
  • Oligopeptides
  • Histidine
  • Cytochrome P-450 Enzyme System