Improvement of proteolytic and oxidative stability of Chondroitinase ABC I by cosolvents.
Recently, utilization of the enzyme Chondroitinase ABC I (cABC I) has received considerable attention in treatment of spinal cord injury. cABC I removes chondroitin sulfate proteoglycans which are inhibitory to axon growth and enhances nerve regeneration. Therefore, determination of cABC I resistance to proteolysis and oxidation provides valuable information for optimizing its clinical application. In this work, proteolytic stability of cABC I to trypsin and chymotrypsin as well as its oxidative resistance to H2O2 was measured. Moreover, the effect of cosolvents glycerol, sorbitol and trehalose on cABC I proteolytic and oxidative stability was determined. The results indicated that cABC I is highly susceptible to proteolysis and oxidation. Comparison of proteolytic patterns demonstrated a high degree of similarity which confirmed the exposure of specific regions of cABC I to proteolysis. However, proteolytic degradation was significantly reduced in the presence of cosolvents. In addition, cosolvents decreased the rate of both cABC I proteolytic and oxidative inactivation. Notably, the degree of stabilization provided by these cosolvents varied greatly. These findings indicated the high potential of cosolvents in protein stabilization to proteolysis and oxidative inactivation.
Department of Clinical Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran. Electronic address: mnazari@kmu.ac.ir.
Mahdieh
M
Nazari-Robati
Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Abolfazl
A
Golestani
Department of Clinical Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
GholamReza
G
Asadikaram
D001426:Bacterial Proteins / Q000737:chemistry*; D019765:Chondroitin ABC Lyase / Q000737:chemistry*; D002918:Chymotrypsin / Q000737:chemistry; D004795:Enzyme Stability; D005990:Glycerol / Q000737:chemistry*; D010084:Oxidation-Reduction; D059748:Proteolysis; D011514:Proteus vulgaris / Q000201:enzymology*; D012997:Solvents / Q000737:chemistry; D013012:Sorbitol / Q000737:chemistry*; D014199:Trehalose / Q000737:chemistry*; D014357:Trypsin / Q000737:chemistry
D001426:Bacterial Proteins; D012997:Solvents; D013012:Sorbitol; D014199:Trehalose; D002918:Chymotrypsin; D014357:Trypsin; D019765:Chondroitin ABC Lyase; D005990:Glycerol
Chondroitinase ABC I; Cosolvents; Limited proteolysis; Oxidative stability; Proteolytic stability