Expression and Isolation of N-Terminal Truncated Human Recombinant Renalase in Prokaryotic Cells

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Published: Sep 30, 2021
DOI: https://doi.org/10.18097/BMCRM00158
Keywords:
isolation; protein solubilization; inclusion bodies; protein expression; renalase

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V.I. Fedchenko
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
A.A. Kaloshin
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
S.A. Kaloshina
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
A.E. Medvedev
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia

Abstract

Renalase (RNLS) is a flavoproteinin which its N-terminal peptide (residues 1-17) has several important functions. In cells, it participates in the formation of the so-called Rossmanfold (residues 2-35), needed for «accommodation» of the FAD cofactor and for performing the catalytic functions of RNLS as a FAD-dependent oxidoreductase (EC 1.6.3.5). RNLS secretion into the extracellular space is accompanied by cleavage of this peptide. The resultant truncated extracellular RNLS cannot bind FAD and therefore performs various noncatalytic functions. In this work, we have performed expression the genetic construct encoding RNLS lacking its N-terminal signal peptide (tRNLS) in E. coli Rosetta (DE3) cells. The recombinant protein was accumulated in inclusion bodies in an insoluble form, which could be solubilized in the presence of a high concentration of urea or guanidine chloride. In contrast to full-length RNLS, which was effectively solubilized in the presence of 8 M urea, tRNLS was preferentially solubilized in the presence of 6 M guanidine chloride.

Article Details

How to Cite
Fedchenko, V., Kaloshin, A., Kaloshina, S., & Medvedev, A. (2021). Expression and Isolation of N-Terminal Truncated Human Recombinant Renalase in Prokaryotic Cells. Biomedical Chemistry: Research and Methods, 4(3), e00158. https://doi.org/10.18097/BMCRM00158
Issue
Vol. 4 No. 3 (2021)
Section
EXPERIMENTAL RESEARCH

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