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Ternary System of Solution Additives with Arginine and Salt for Refolding of Beta-Galactosidase

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Abstract

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 (β-gal) using Arg HCl and other salts. In a binary system using only Arg HCl, the refolding yield of β-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.

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Abbreviations

Arg HCl:

l-Arginine hydrochloride

β-gal:

Beta-galactosidase

DTT:

Dithiothreitol

EDTA:

Ethylenediaminetetraacetic acid

Gdn HCl:

Guanidine hydrochloride

Tween 20:

Polyoxyethylene sorbitan monolaurate

Na3-citrate:

Trisodium citrate

Tris-HCl:

Tris(hydroxymethyl)aminomethane hydrochloride

ONPG:

o-Nitrophenyl-β-d-galactopyranoside

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Acknowledgments

This work was partly supported by the Tsukuba Industrial Liaison and Cooperative Research Center.

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Authors and Affiliations

  1. Institute of Applied Physics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8573, Japan

    Akiko Fujimoto, Atsushi Hirano & Kentaro Shiraki

Authors
  1. Akiko Fujimoto
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  2. Atsushi Hirano
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  3. Kentaro Shiraki
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Correspondence to Kentaro Shiraki.

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Fujimoto, A., Hirano, A. & Shiraki, K. Ternary System of Solution Additives with Arginine and Salt for Refolding of Beta-Galactosidase. Protein J 29, 161–166 (2010). https://doi.org/10.1007/s10930-010-9235-7

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  • DOI: https://doi.org/10.1007/s10930-010-9235-7

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