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Dynamic Light-scattering Analysis Of The Electrostatic Interaction Of Hexahistidine-tagged Cytochrome P450 Enzyme With Semiconductor Quantum Dots.

Binil Itty Ipe, Anuj Shukla, Huachang Lu, Bo Zou, Heinz Rehage, Christof M. Niemeyer
Published 2006 · Chemistry, Medicine
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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.



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