Hostname: page-component-586b7cd67f-g8jcs Total loading time: 0 Render date: 2024-11-25T13:13:02.317Z Has data issue: false hasContentIssue false
  • English
  • Français

Functional characterization of X-prolyl aminopeptidase from Toxoplasma gondii

Published online by Cambridge University Press:  25 May 2016

MINGFA YANG ,
JUN ZHENG ,
HONGLIN JIA  and
MINGXIN SONG
MINGFA YANG
Affiliation:
College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, China Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Harbin, China
JUN ZHENG
Affiliation:
Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Harbin, China
HONGLIN JIA*
Affiliation:
Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Harbin, China
MINGXIN SONG*
Affiliation:
College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, China
*
*Corresponding authors: College of Veterinary Medicine, Northeast Agricultural University, Mucai Street 59, Xiangfang District, Harbin 150001, People's Republic of China. Tel: +8613059000733. E-mail: [email protected] and Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Maduan Street 427, Nangang District, Harbin 150001, People's Republic of China. Tel: +8618946066070. E-mail: [email protected]
*Corresponding authors: College of Veterinary Medicine, Northeast Agricultural University, Mucai Street 59, Xiangfang District, Harbin 150001, People's Republic of China. Tel: +8613059000733. E-mail: [email protected] and Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Maduan Street 427, Nangang District, Harbin 150001, People's Republic of China. Tel: +8618946066070. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

In the present study, a recombinant aminopeptidase P (rTgAPP) from Toxoplasma gondii was expressed in Escherichia coli to evaluate its enzyme parameters. The rTgAPP showed strong activity against a synthetic substrate for aminopeptidase P at pH 8·0 with a Km value of 0·255 µm and a kcat value of 35·6 s−1. The overall catalytic efficiency (kcat/Km) of the rTgAPP was 139·6 × 105 M−1 s−1. The activity of rTgAPP was enhanced by the addition of divalent cations and inhibited by bestatin. Deletion of TgAPP gene in the parasite through a CRISPR/Cas9 system resulted in inhibition of growth indicating the importance of TgAPP. Thus our findings reveal that TgAPP is an active enzyme in T. gondii and provide an insight into the function of TgAPP.

Keywords

Toxoplasma gondiiToxoplasmosisX-prolyl aminopeptidaseenzymatic activitygrowth
Type
Research Article
Information
Parasitology , Volume 143 , Issue 11 , September 2016 , pp. 1443 - 1449
Check for updates
Copyright
Copyright © Cambridge University Press 2016 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Abe, F., Alvord, G., Koyama, M., Matsuda, A. and Talmadge, J. E. (1989). Pharmacokinetics of bestatin and oral activity for treatment of experimental metastases. Cancer Immunology Immunotherapy 28, 29–33.CrossRefGoogle ScholarPubMed
Ajioka, J. W. and Soldati, D. (2007). Molecular and cellular biology. Veterinary Parasitology 153, 195.Google Scholar
Centers for Disease Control and Prevention (2015). Toxoplasmosis. Parasites. Centers for Disease Control and Prevention, Atlanta, USA.Google Scholar
Cunningham, D. F. and O'Connor, B. (1997). Proline specific peptidases. Biochimica et Biophysica Acta 1343, 160–186.Google Scholar
Dalal, S. and Klemba, M. (2007). Roles for two aminopeptidases in vacuolar hemoglobin catabolism in Plasmodium falciparum . Journal of Biological Chemistry, 282, 35978–35987.Google Scholar
Dou, Z., McGovern, O. L., Cristina, M. D. and Carruthers, V. B. (2014). Toxoplasma gondii ingests and digests host cytosolic proteins. MBio 5, e01188–14.Google Scholar
Hooper, N. M., Hryszko, J. and Turner, A. J. (1990). Purification and characterization of pig kidney aminopeptidase P. Journal of Biological Chemistry 267, 509–515.Google Scholar
Lee, H. S., Kim, Y., Lee, J. H. and Bae, S. S. (2006). Cloning, expression and characterization of aminopeptidase P from the Hyperthermophilic Archaeon Thermococcus sp. Strain NA1. Applied and Environmental Microbiology 72, 1886–1890.Google Scholar
Mars, I. and Monnet, V. (1995). An aminopeptidase P from Lactococcus lactis with original specificity. Biochimica et Biophysica Acta 1243, 209–215.CrossRefGoogle ScholarPubMed
Phang, J. M., Liu, W. and Zabirnyk, O. (2010). Proline metabolism and microenvironmental stress. Annual Review of Nutrition 30, 441–463.Google Scholar
Ragheb, D., Bompiani, D., Dalal, S. and Klembal, M. (2009). Evidence for catalytic roles for Plasmodium falciparum aminopeptidase P in the food vacuole and cytosol. Journal of Biological Chemistry 284, 24806–24815.Google Scholar
Rosenthal, P. J. (2004). Cysteine proteases of malaria. International Journal for Para-sitology 34, 1489–1499.Google Scholar
Rusu, I. and Yaron, A. (1992). Aminopeptidase P from human leukocytes. European Journal of Biochemistry, 210, 93–100.Google Scholar
Schorlemmer, H. U., Bosslet, K. and Sedlacek, H. H. (1983). Ability of the immunomodulating dipeptide bestatin to activate cytotoxic mononuclear phagocytes. Cancer Research 43, 4148–4153.Google ScholarPubMed
Shen, B., Brown, K. M., Lee, T. D. and Sibley, L. D. (2014). Efficient gene disruption in diverse strains of Toxoplasma gondii using CRISPR/CAS9. MBio 5, e01114–14.Google Scholar
Taylor, A. (1993). Aminopeptidases: structure and function. FASEB Journal 7, 290–298.Google Scholar
Torda, A. (2001). Toxoplasmosis. Are cats really the source? Australian Family Physician 30, 743–747.Google Scholar
Turner, A. J., Hyde, R. J., Lim, J. and Hooper, N. M. (1997). Structural studies of aminopeptidase P. A novel cellular peptidase. Advances in Experimental Medicine and Biology 421, 7–16.Google Scholar
Viglio, S., Annovazzi, L., Conti, B., Genta, I., Perugini, P., Zanone, C., Casado, B., Cetta, G. et al. (2006). The role of emerging techniques in the investigation of prolidase deficiency: from diagnosis to the development of a possible therapeutical approach. Journal of Chromatography B 832, 1–8.Google Scholar
Zheng, J., Jia, H. and Zheng, Y. (2014). Knockout of leucine aminopeptidase in Toxoplasma gondii using CRISPR/Cas9. International Journal for Parasitology 45, 141–148.Google Scholar
Supplementary material: Image

Yang supplementary material

Figure S1

Download Yang supplementary material(Image)
Image 4.2 MB
Supplementary material: Image

Yang supplementary material

Figure S2

Download Yang supplementary material(Image)
Image 5.2 MB
Supplementary material: Image

Yang supplementary material

Figure S3

Download Yang supplementary material(Image)
Image 1.6 MB
Supplementary material: File

Yang supplementary material

Yang supplementary material 1

Download Yang supplementary material(File)
File 14 KB