L-Arginine hydrochloride increases the solubility of folded and unfolded recombinant plasminogen activator rPA.

18:25 EDT 23rd September 2014 | BioPortfolio

Summary of "L-Arginine hydrochloride increases the solubility of folded and unfolded recombinant plasminogen activator rPA."

L-Arginine hydrochloride (L-ArgHCl) was found to be an effective enhancer for in vitro protein refolding more than two decades ago. A detailed understanding of the mechanism of action, by which L-ArgHCl as co-solvent is capable to effectively suppress protein aggregation, while protein stability is preserved, has remained elusive. Concepts for the effects of co-solvents, which have been established over the last decades, were found to be insufficient to completely explain the effects of L-ArgHCl on protein refolding.In this paper we present data, which clearly establish that L-ArgHCl acts on the equilibrium solubility of the native model protein rPA, while for S-carboxymethylated rPA (IAA-rPA) that served as a model protein for denatured protein states, equilibrium solubilities could not be obtained. Solid to solute free transfer energies for native rPA were lowered by up to 14 kJ mol(-1) under the tested conditions. This finding is in marked contrast to a previously proposed model in which L-ArgHCl acts as a neutral crowder which exclusively has an influence on the stability of the transition state of aggregation. The effects on the apparent solubility of IAA-rPA as well as on the aggregation kinetics of all studied protein species that were observed in the present work could tentatively explained within the framework of a nucleation-aggregation scheme, in which L-ArgHCl exerts a strong effect on the pre-equilibria leading to formation of the aggregation seed.

Affiliation

Institut für Biochemie und Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany.

Journal Details

This article was published in the following journal.

Name: Protein science : a publication of the Protein Society
ISSN: 1469-896X
Pages:

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Medical and Biotech [MESH] Definitions

A proteolytic enzyme that converts PLASMINOGEN to FIBRINOLYSIN where the preferential cleavage is between ARGININE and VALINE. It was isolated originally from human URINE, but is found in most tissues of most VERTEBRATES.

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Conformational transitions of a protein from unfolded states to a more folded state.

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An extracellular receptor specific for UROKINASE-TYPE PLASMINOGEN ACTIVATOR. It is attached to the cell membrane via a GLYCOSYLPHOSPHATIDYLINOSITOL LINKAGE and plays a role in the co-localization of urokinase-type plasminogen activator with PLASMINOGEN.

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