Next Article in Journal
Doxycycline Alters the Porcine Renal Proteome and Degradome during Hypothermic Machine Perfusion
Previous Article in Journal
Resveratrol Downregulates Granulocyte-Macrophage Colony-Stimulating Factor-Induced Oncostatin M Production through Blocking of PI3K/Akt/NF-κB Signal Cascade in Neutrophil-like Differentiated HL-60 Cells
 
 
Article

The Small Metal-Binding Protein SmbP Improves the Expression and Purification of the Recombinant Antitumor-Analgesic Peptide from the Chinese Scorpion Buthus martensii Karsch in Escherichia coli

1
Facultad de Ciencias Quimicas, Universidad Autonoma de Nuevo Leon, Av. Universidad s/n, Cd. Universitaria, San Nicolas de los Garza 66455, Mexico
2
Departamento de Patologia Clinica, Hospital Universitario Dr. Jose Eleuterio Gonzalez, Universidad Autonoma de Nuevo Leon, Monterrey 64460, Mexico
*
Author to whom correspondence should be addressed.
Academic Editor: Julius Liobikas
Curr. Issues Mol. Biol. 2022, 44(2), 550-558; https://doi.org/10.3390/cimb44020038
Received: 3 December 2021 / Revised: 14 January 2022 / Accepted: 18 January 2022 / Published: 22 January 2022
(This article belongs to the Section Molecular Medicine)
We have recently shown that SmbP, the small metal-binding protein of Nitrosomonas europaea, can be employed as a fusion protein to express and purify recombinant proteins and peptides in Escherichia coli. SmbP increases solubility, allows simple, one-step purification through affinity chromatography, and provides superior final yields due to its low molecular weight. In this work, we report for the first time the use of SmbP to produce a recombinant peptide with anticancer activity: the antitumor-analgesic peptide (BmK-AGAP), a neurotoxin isolated from the venom of the Chinese scorpion Buthus martensii Karsch. This peptide was expressed in Escherichia coli SHuffle for correct, cytoplasmic, disulfide bond formation and tagged with SmbP at the N-terminus to improve its solubility and allow purification using immobilized metal affinity chromatography. SmbP_BmK-AGAP was found in the soluble fraction of the cell lysate. After purification and removal of SmbP by digestion with enterokinase, 1.8 mg of pure and highly active rBmK-AGAP was obtained per liter of cell culture. rBmK-AGAP exhibited antiproliferative activity on the MCF-7 cancer cell line, with a half-maximal inhibitory concentration value of 7.24 μM. Based on these results, we considered SmbP to be a suitable carrier protein for the production of recombinant, biologically active BmK-AGAP. We propose that SmbP should be an attractive fusion protein for the expression and purification of additional recombinant proteins or peptides that display anticancer activities. View Full-Text
Keywords: SmbP; small metal-binding protein; BmK-AGAP; Escherichia coli; recombinant peptides; anticancer activity SmbP; small metal-binding protein; BmK-AGAP; Escherichia coli; recombinant peptides; anticancer activity
Show Figures

Figure 1

MDPI and ACS Style

Martinez-Mora, E.; Arredondo-Espinoza, E.; Casillas-Vega, N.G.; Cantu-Cardenas, M.E.; Balderas-Renteria, I.; Zarate, X. The Small Metal-Binding Protein SmbP Improves the Expression and Purification of the Recombinant Antitumor-Analgesic Peptide from the Chinese Scorpion Buthus martensii Karsch in Escherichia coli. Curr. Issues Mol. Biol. 2022, 44, 550-558. https://doi.org/10.3390/cimb44020038

AMA Style

Martinez-Mora E, Arredondo-Espinoza E, Casillas-Vega NG, Cantu-Cardenas ME, Balderas-Renteria I, Zarate X. The Small Metal-Binding Protein SmbP Improves the Expression and Purification of the Recombinant Antitumor-Analgesic Peptide from the Chinese Scorpion Buthus martensii Karsch in Escherichia coli. Current Issues in Molecular Biology. 2022; 44(2):550-558. https://doi.org/10.3390/cimb44020038

Chicago/Turabian Style

Martinez-Mora, Evelyn, Eder Arredondo-Espinoza, Nestor G. Casillas-Vega, Maria Elena Cantu-Cardenas, Isaias Balderas-Renteria, and Xristo Zarate. 2022. "The Small Metal-Binding Protein SmbP Improves the Expression and Purification of the Recombinant Antitumor-Analgesic Peptide from the Chinese Scorpion Buthus martensii Karsch in Escherichia coli" Current Issues in Molecular Biology 44, no. 2: 550-558. https://doi.org/10.3390/cimb44020038

Find Other Styles

Article Access Map by Country/Region

1
Back to TopTop