A new artificial chaperone for protein refolding: sequential use of detergent and alginate.
A novel artificial chaperone system using a combination of detergents and alginate was developed to refold three enzymes with totally different structures. Upon dilution of denatured protein in the presence of the capturing agent, complexes of the detergent and non-native protein molecules are formed and thereby the formation of protein aggregates is prevented. The so-called captured protein is unable to refold from the detergent-protein complex states unless a stripping agent is used to gradually remove the detergent molecules. In that respect, we used alginate, a linear copolymer of D-mannuronic acid and L-guluronic acid, to initiate and complete the refolding process. The results indicated that the extent of refolding assistance for the proteins was different due to detergent structure and also the length of hydrophobic portion of each detergent. These observed differences were attributed to the strong electrostatic and hydrophobic interactions among the capturing and stripping agents used in this investigation. Based on this newly developed method, it is expected that the protein refolding operation can be achieved easily, cheaply and efficiently.
Khodagholi
Fariba
F
Institute of Biochemistry and Biophysics, The University of Tehran, P.O. Box 13145-1384, Tehran, Iran.
Eftekharzadeh
Bahareh
B
Yazdanparast
Razieh
R
D000464:Alginates / Q000737:chemistry*; D000469:Alkaline Phosphatase / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D003902:Detergents / Q000737:chemistry*; D019855:Ethylene Glycol / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D018832:Molecular Chaperones / Q000737:chemistry*; D017510:Protein Folding; D012492:Salts / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D000516:alpha-Amylases / Q000737:chemistry
D000464:Alginates; D003902:Detergents; D006603:Hexuronic Acids; D018832:Molecular Chaperones; D012492:Salts; D020723:Glucuronic Acid; D000469:Alkaline Phosphatase; D000516:alpha-Amylases; D002256:Carbonic Anhydrases; D019855:Ethylene Glycol