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Interactions protéines-nanoparticules : émergence de nouveaux facteurs déterminant la formation de la couronne de protéines
Laurent Marichal
Fri, Apr. 27th 2018, 00:00
Amphi. Bloch, Bât. 774, Orme des Merisiers, CEA-Saclay

Manuscrit de la thèse.


Résumé :

Nanoparticles are ubiquitous in our environment and their presence inside our bodies is now established. Besides, in a biological medium, nanoparticles are spontaneously covered by proteins that form the so-called protein corona. Depending on the corona composition, a nanoparticle will possess a specific "biological identity" conditioning its biodistribution as well as its potential toxicity.Despite being highly studied, many aspects of the protein adsorption mechanisms remain unknown. Here we particularly focused on the influence of two physicochemical characteristics, which had rarely been addressed: protein size and post-translational modifications. Also, because of their intensive use, we worked on silica nanoparticles (SiNPs).We studied the adsorption of hemoproteins on SiNPs, both of them having different sizes. Adsorption isotherms and calorimetry studies showed a relationship between the protein size and its affinity towards silica surfaces. Finer differences could also be observed by varying the SiNPs size. Additionally, structural analyses of adsorbed proteins were performed using circular dichroism and small-angle neutron scattering. The adsorption of hemoproteins, which are well-structured proteins, seems to have little effects on their structure. However, even though the quaternary structure is maintained, structural modifications can be seen.Using yeast protein extracts and synthetic peptides, the major role of arginine asymmetric dimethylation on proteins/SiNPs interaction could be established. The use of experimental and simulation techniques allowed us to understand the mechanism responsible for the high affinity of peptides having this peculiar methylation. As a whole, this work suggests that post-translational modifications can influence considerably the interactions between biomolecules and mineral surfaces.

Mots-clés : Nanoparticule, Silice, Protéine, Couronne


Proteins-nanoparticles interactions : influence of emerging factors affecting the protein corona formation

Abstract:

Nanoparticles are ubiquitous in our environment and their presence inside our bodies is now established. Besides, in a biological medium, nanoparticles are spontaneously covered by proteins that form the so-called protein corona. Depending on the corona composition, a nanoparticle will possess a specific "biological identity" conditioning its biodistribution as well as its potential toxicity.Despite being highly studied, many aspects of the protein adsorption mechanisms remain unknown. Here we particularly focused on the influence of two physicochemical characteristics, which had rarely been addressed: protein size and post-translational modifications. Also, because of their intensive use, we worked on silica nanoparticles (SiNPs).We studied the adsorption of hemoproteins on SiNPs, both of them having different sizes. Adsorption isotherms and calorimetry studies showed a relationship between the protein size and its affinity towards silica surfaces. Finer differences could also be observed by varying the SiNPs size. Additionally, structural analyses of adsorbed proteins were performed using circular dichroism and small-angle neutron scattering. The adsorption of hemoproteins, which are well-structured proteins, seems to have little effects on their structure. However, even though the quaternary structure is maintained, structural modifications can be seen.Using yeast protein extracts and synthetic peptides, the major role of arginine asymmetric dimethylation on proteins/SiNPs interaction could be established. The use of experimental and simulation techniques allowed us to understand the mechanism responsible for the high affinity of peptides having this peculiar methylation. As a whole, this work suggests that post-translational modifications can influence considerably the interactions between biomolecules and mineral surfaces.

Keywords: Nanoparticle, Silica, Protein, Adsorption, Interaction, Corona

Contact : Serge PIN

 

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