Overexpression, purification, molecular characterization and pharmacological evaluation for anticancer activity of ribosomal protein S23 from the giant panda (Ailuropoda melanoleuca)

Wang,Ting; Hou,Yiling; Ding,Xiang; Song,Bo; Wang,Fang; Hou,Wanru

doi:10.3892/mmr.2013.1430

June 2013 Volume 7 Issue 6

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

June 2013 Volume 7 Issue 6

Full Size Image

Article

Overexpression, purification, molecular characterization and pharmacological evaluation for anticancer activity of ribosomal protein S23 from the giant panda (Ailuropoda melanoleuca)

Authors:
- Ting Wang
- Yiling Hou
- Xiang Ding
- Bo Song
- Fang Wang
- Wanru Hou
View Affiliations / Copyright

Affiliations: Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education), College of Life Sciences, China West Normal University, Nanchong, Sichuan 637009, P.R. China
Pages: 1875-1882
|
Published online on: April 16, 2013

https://doi.org/10.3892/mmr.2013.1430
Expand metrics +

Abstract

Ribosomal protein S23 (RPS23) is a component of the 40S small ribosomal subunit encoded by the RPS23 gene, which is specific to eukaryotes. The cDNA and genomic sequence of RPS23 were cloned from Ailuropoda melanoleuca (A. melanoleuca) using reverse transcription‑polymerase chain reaction (RT-PCR) technology and touchdown PCR, respectively. The two sequences were analyzed preliminarily and the cDNA of the RPS23 gene was overexpressed in Escherichia coli (E. coli) BL21. The cDNA of RPS23 cloned from giant panda was 472 bp, and it contained an open reading frame (ORF) of 432 bp encoding 142 amino acids. The nucleotide sequence of the coding sequence showed a high degree of homology to some mammals as determined by BLAST analysis, similar to the amino acid sequence. The genomic sequence was 2,105 bp in length, with 4 exons and 3 introns. The primary structure analysis revealed that the molecular weight of the putative RPS23 protein was 15.80 kDa with a theoretical isoelectric point (pI) of 11.23. The molecular weight of the recombinant protein RPS23 was 21.5 kDa with a theoretical pI of 10.57. Topology prediction showed that there are seven different patterns of functional sites in the RPS23 protein of giant panda. RPS23 was successfully expressed in E. coli and its protein fused with the N‑terminal His‑tagged protein triggered the accumulation of an expected 21.5‑kDa polypeptide. The inhibitory rate of tumor growth in mice treated with 0.1 µg/ml RPS23 protein was 49.45%, the highest in the three doses used, which may be comparable to mannatide treatment. Histology of immune organs showed that the tissues were characterized by a regular and tight arrangement, while tumor tissues of the mice in the RPS23 group exhibited a loose arrangement compared to the control group. However, there was no obvious damage to other organs, such as the heart, lung and kidney. Investigations are currently being conducted to determine the bioactive principles of the recombinant protein RPS23 responsible for its anticancer activity.

View Figures

View References

1	Wool IG: Extraribosomal functions of ribosomal proteins. Trends Biochem Sci. 21:164–165. 1996. View Article : Google Scholar : PubMed/NCBI
2	Wool IG, Chan YL and Glück A: Structure and evolution of mammalian ribosomal proteins. Biochem Cell Biol. 73:933–947. 1995. View Article : Google Scholar : PubMed/NCBI
3	Fan W, Christensen M, Eichler E, Zhang X and Lennon G: Cloning, sequencing, gene organization, and localization of the human ribosomal protein RPL23A gene. Genomics. 46:234–239. 1997. View Article : Google Scholar : PubMed/NCBI
4	Yang F and Liu WP: The progress of ribosomal protein genes and human diseases. J Clin Exp Pathol. 20:354–356. 2005.
5	Elantak L, Wagner S, Herrmannová A, Karásková M, Rutkai E, Lukavsky PJ and Valásek L: The indispensable N-terminal half of eIF3j/HCR1 cooperates with its structurally conserved binding partner eIF3b/PRT1-RRM and with eIF1A in stringent AUG selection. J Mol Biol. 396:1097–1116. 2010. View Article : Google Scholar : PubMed/NCBI
6	Unbehaun A, Marintchev A, Lomakin IB, Didenko T, Wagner G, Hellen CU and Pestova TV: Position of eukaryotic initiation factor eIF5B on the 80S ribosome mapped by directed hydroxyl radical probing. EMBO J. 26:3109–3123. 2007. View Article : Google Scholar : PubMed/NCBI
7	Huang X, Chen Y, Li WB, Cohen SN, Liao FF, Li L, Xu H and Zhang YW: The Rps23rg gene family originated through retroposition of the ribosomal protein s23 mRNA and encodes proteins that decrease Alzheimer’s beta-amyloid level and tau phosphorylation. Hum Mol Genet. 19:3835–3843. 2010.PubMed/NCBI
8	Anand P, Kunnumakkara AB, Sundaram C, Harikumar KB, Tharakan ST, Lai OS, Sung B and Aggarwal BB: Cancer is a preventable disease that requires major lifestyle changes. Pharm Res. 25:2097–2116. 2008. View Article : Google Scholar : PubMed/NCBI
9	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
10	Tolar J and Neglia JP: Transplacental and other routes of cancer transmission between individuals. J Pediatr Hematol Oncol. 25:430–434. 2003. View Article : Google Scholar : PubMed/NCBI
11	Croce CM: Oncogenes and cancer. N Engl J Med. 358:502–511. 2008. View Article : Google Scholar
12	Wu GF, Hou YL, Hou WR, Song Y and Zhang T: Giant pandaribosomal protein s14: cDNA, genomic sequence cloning, sequence analysis, and overexpression. Genet Mol Res. 9:2004–2015. 2010. View Article : Google Scholar : PubMed/NCBI
13	Du YJ, Luo XY, Hao YZ, Zhang T and Hou WR: Cloning and overexpression of acidic ribosomal phosphoprotein P1 gene (RPLP1) from the giant panda. Inter J Biol Sci. 3:428–433. 2007.PubMed/NCBI
14	Hou WR, Chen Y, Peng ZS, Wu X and Tang ZX: cDNA cloning and sequences analysis of ubiquinol-cytochrome c reductase complex ubiquinone-binding protein (QP-C) from giant panda. Acta Theriol Sin. 27:190–194. 2007.
15	Hou YL, Du YJ, Hou WR, Zhou CQ, Hao YZ and Zhang T: Cloning and sequence analysis of translocase of inner mitochondrial membrane 10 homolog (yeast) gene (TIMM10) from the giant panda. J Cell Anim Biol. 3:9–14. 2009.
16	Liao MJ, Zhu MY, Zhang ZH, Zhang AJ, Li GH and Sheng FJ: Cloning and sequence analysis of FSH and LH in the giant panda (Ailuropoda melanoleuca). Anim Reprod Sci. 77:107–116. 2003. View Article : Google Scholar : PubMed/NCBI
17	Wu ZA, Liu WX, Murphy C and Gall J: Satellite 1 DNA sequence from genomic DNA of the giant panda Ailuropoda melanoleuca. Nucleic Acids Res. 18:10541990. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Overexpression, purification, molecular characterization and pharmacological evaluation for anticancer activity of ribosomal protein S23 from the giant panda (Ailuropoda melanoleuca)

This article is mentioned in:

Abstract

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Related Articles