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Protein disulphide isomerase-induced refolding of sonochemically prepared Ribonuclease A microspheres

Paper ID Volume ID Publish Year Pages File Format Full-Text
23695 43463 2012 5 PDF Available
Title
Protein disulphide isomerase-induced refolding of sonochemically prepared Ribonuclease A microspheres
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.

► Sonochemically prepared RNase A microspheres. ► Changes on the physicochemical conformational and biological properties of RNase A after microspherization. ► Protein disulphide isomerase with oxidized and reduced active site application on microspheres. ► Increased biological activity and protein release after enzyme application.

Keywords
Protein disulphide isomerase; Ribonuclease A; Ultrasound; Microspheres; Activity recovery; Protein release; Protein folding
First Page Preview
Protein disulphide isomerase-induced refolding of sonochemically prepared Ribonuclease A microspheres
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 159, Issues 1–2, 31 May 2012, Pages 78–82
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering