Using magnetic nanoparticles to probe protein damage in ferritin caused by freeze concentration

Autor: E. F. Chagas, S. Correia Carreira, W. Schwarzacher
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Zdroj: AIP Advances, Vol 5, Iss 11, Pp 117201-117201-5 (2015)
Druh dokumentu: article
ISSN: 2158-3226
DOI: 10.1063/1.4935261
Popis: We demonstrate a method for monitoring the damage caused to a protein during freeze-thawing in the presence of glycerol, a cryo-protectant. For this work we synthesized magnetite nanoparticles doped with 2.5% cobalt inside the protein ferritin (CMF), dissolved them in different concentration glycerol solutions and measured their magnetization after freezing in a high applied field (5 T). As the temperature was raised, a step-like decrease in the sample magnetization was observed, corresponding to the onset of Brownian relaxation as the viscosity of the freeze-concentrated glycerol solution decreased. The position of the step reveals changes to the protein hydrodynamic radius that we attribute to protein unfolding, while its height depends on how much protein is trapped by ice during freeze concentration. Changes to the protein hydrodynamic radius are confirmed by dynamic light scattering (DLS) measurements, but unlike DLS, the magnetic measurements can provide hydrodynamic data while the solution remains mainly frozen.
Databáze: Directory of Open Access Journals