Impact of ConcanavalinA affinity in the intracellular fate of Protein Corona on Glucosamine Au nanoparticles

Desirè Di Silvio,Laura Polito,Sergio E Moya,Luigi Lay,Alessandro Silvestri

doi:10.1038/s41598-018-27418-w

Abstract

Biological fate and toxicity of nanoparticles (NPs) are connected to the interaction between NPs and the protein corona (PC) spontaneously forming around NPs in biological matrixes. PC is a dynamic entity that confers biological identity to NPs. In this work, fluorescence cross-correlation spectroscopy (FCCS) is used to study the impact of specific interactions between the NP surface and proteins on the intracellular fate of PC. The stability of the PC formed around glucosamide-functionalized Au-NPs from ConcanavalinA (ConA) or Bovine Serum Albumin (BSA) is characterized by FCCS. The NPs show higher affinity for ConA and competitive assays show that ConA easily exchanges BSA. A549 cells are exposed to glucosamide-functionalized Au-NPs with preformed ConA and BSA PCs. Intracellularly the frequency of cross-correlation for Au NPs with ConA PC remains constant to a 70% value until 24 h while for BSA it decreases to a 15% during the same period. FCCS measurements in several locations in the cell point out a different level of aggregation for the NPs with either ConA or BSA PCs. Our results show that the affinity of NPs functionalized with a ligand with affinity for a specific protein in bulk is retained intracellularly influencing NP fate and translocation.

Highlights

The surface chemistry of nanoparticles (NPs) plays a fundamental role on their interaction with biomolecules and their fate in biological matrixes
We have studied by means of Fluorescence correlation spectroscopy (FCS) technique is the cross-correlation spectroscopy (FCCS), both in vitro and in the live cell, the strength of the interaction between the glucosamine Au NPs and ConA
We compared it with the affinity of the NPs for bovine serum albumin (BSA), which has nonspecific interactions with glucosamide, besides being the most abundant protein in plasma and it can as well be taken as a model for the plasma itself

Summary

Corona on Glucosamine Au

Received: 3 January 2018 Accepted: 25 May 2018 Published: xx xx xxxx nanoparticles. Desirè Di Silvio[1], Alessandro Silvestri[2,3,6], Luigi Lay[3,4], Laura Polito5 & Sergio E. PC is a dynamic entity that confers biological identity to NPs. In this work, fluorescence cross-correlation spectroscopy (FCCS) is used to study the impact of specific interactions between the NP surface and proteins on the intracellular fate of PC. We apply FCCS for the first time to study the intracellular stability of the PC when it is formed from a protein with a specific affinity for the NP coating. We have studied by means of FCCS, both in vitro and in the live cell, the strength of the interaction between the glucosamine Au NPs and ConA We compared it with the affinity of the NPs for bovine serum albumin (BSA), which has nonspecific interactions with glucosamide, besides being the most abundant protein in plasma and it can as well be taken as a model for the plasma itself. The degradation of the PC will result in the NP surface directly exposed to other biomolecules with potential impact on cell metabolism and toxicity[39,40]

Results and Discussion

PC BSAd

Conclusions

Additional Information