Protein disulphide isomerase-induced refolding of sonochemically prepared Ribonuclease A microspheres |
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Authors: | Fernandes Margarida M Silva Raquel Ferreira Helena Donelli Ilaria Freddi Giuliano Cavaco-Paulo Artur |
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Affiliation: | Textile Engineering Department, University of Minho, Campus of Azurém, Guimar?es, Portugal. |
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Abstract: | The present communication describes for the first time the development of Ribonuclease A (RNase A) microspheres using the sonochemical method followed by an enzymatic treatment with protein disulphide isomerase (PDI). Ultrasound application induced changes on the protein physicochemical and biological properties: the enzymatic activity of RNase A was decreased in 35% and the free thiol groups content was significantly increased, probably due to the breakage of protein disulphide bonds and assembly of RNase A monomers. The deconvolution of amide I band, from Fourier Transform Infrared Spectroscopy, showed that the secondary structure of RNase A was slightly changed after microspherization. The PDI application on microspheres promoted the recovery of RNase A biological activity and induced the release of active protein into solution in its native state. These results were promoted by different states of PDI active site: oxidized and reduced, respectively. The PDI aptitude to catalyze the refolding of a protein substrate in the form of spheres is here reported. |
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