A method to measure the denatured proteins in the corona of nanoparticles based on the specific adsorption of Hsp90ab1.

Nanoscale
Shang LiuLei Dong

Abstract

The protein corona influences and determines the biological function of nanoparticles (NPs) in vivo. Analysis and understanding of the activities of proteins in coronas are crucial for nanobiology and nanomedicine research. Misfolded proteins in the corona of NPs theoretically exist, and a protein denaturation-related cellular response might occur in this process as well as in related diseases. The exact evaluation of protein denaturation in the corona is valuable to assess the bioactivities of NPs. Here, we found that the level of adsorbed heat shock protein 90 kDa α class B member 1 (Hsp90ab1) by the denatured protein in iron-cobalt-nickel alloy NPs (FeCoNi NPs) and iron oxide NPs (Fe3O4 NPs) was correlated with circular dichroism (CD) analysis and 1-anilinonaphthalene-8-sulfonate (ANS) analysis. The content of Hsp90ab1 in the corona could be easily analysed by western blotting (WB). Further analysis suggested that the method could precisely show the time-dependent protein denaturation on Fe3O4 NPs, as well as the influence of the size and the surface modification. More importantly, this method could be applied to other proteins, like lysozyme, other than albumin. Based on the results and the correlation analysis, incubation ...Continue Reading

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Methods Mentioned

BETA
circular dichroism
transmission electron microscopy
microscale thermophoresis
isothermal titration calorimetry

Software Mentioned

K2D3
Prism
ImageJ
GraphPad
NT Analysis

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