Ternary system of solution additives with arginine and salt for refolding of beta-galactosidase.
(L)-Arginine hydrochloride (Arg HCl) has been used for protein refolding as a universal aggregation suppressor for monomeric proteins. This paper presents an investigation of the refolding of tetrameric beta-galactosidase (beta-gal) using Arg HCl and other salts. In a binary system using only Arg HCl, the refolding yield of beta-gal increased with increasing concentration up to 0.2 M. However, the refolding yield sharply decreased above this concentration, reaching the level below the control yield of 5% at 0.5 M and near zero above 0.75 M, an observation unexpected from monomeric proteins. In a ternary system using both 0.2 M Arg HCl and another salt, the refolding yield increased up to 1.5-fold higher than that in the binary system. These data indicate that aggregation suppressive effects of protein increase with Arg HCl concentration, but also are deleterious to self-association of the protein. This dual nature of Arg HCl effects may have to be taken into account in its application for refolding of oligomeric proteins.
Fujimoto
Akiko
A
Institute of Applied Physics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8573, Japan.
Hirano
Atsushi
A
Shiraki
Kentaro
K
D001120:Arginine / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006851:Hydrochloric Acid / Q000737:chemistry; D017510:Protein Folding; D001616:beta-Galactosidase / Q000737:chemistry* / Q000378:metabolism
D029968:Escherichia coli Proteins; D001120:Arginine; D001616:beta-Galactosidase; D006851:Hydrochloric Acid