Hostname: page-component-586b7cd67f-dsjbd Total loading time: 0 Render date: 2024-11-20T07:32:12.772Z Has data issue: false hasContentIssue false
  • English
  • Français

Molecular characterization and tissue localization of glutathione S-transferase from adult Ancylostoma ceylanicum

Published online by Cambridge University Press:  21 January 2020

J.X. Hang ,
L. He ,
A.M.I. Abuzeid ,
Y. Huang ,
Y.Q. Liu ,
X.X. Yan ,
Q. Zhao ,
X. Li ,
J.M. Liu  and
G.Q. Li Open the ORCID record for G.Q. Li [Opens in a new window]
J.X. Hang*
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
L. He
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
A.M.I. Abuzeid
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
Y. Huang
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
Y.Q. Liu
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
X.X. Yan
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
Q. Zhao
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
X. Li
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
J.M. Liu
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
G.Q. Li
Affiliation:
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou510642, China
*
Author for correspondence: G.Q. Li, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Glutathione S-transferases (GSTs) are a detoxifying enzyme family that is essential for parasite blood-feeding and survival, and represent potential targets for hookworm vaccine development. Multiple GST-encoding complementary DNAs (cDNAs) have been cloned from Ancylostoma caninum and Necator americanus, but there are no reports about the cloning of this enzyme from Ancylostoma ceylanicum, the animal-derived zoonotic hookworm. To study the molecular nature and tissue localization of GST of A. ceylanicum (Ace-GST), we designed primers based on the GST gene sequence of A. ceylanicum in GenBank, amplified the Ace-GST cDNA by reverse transcription polymerase chain reaction, and analysed its homology and genetic evolution relationship. The amplified product was cloned into the pET-32a vector and transformed into Escherichia coli BL21 (DE3) for expression. To prepare anti-GST polyclonal antibodies, the recombinant protein was purified and used to immunize Kunming mice. The level of immunoglobulin G (IgG) antibody in the serum of immunized mice was detected by indirect enzyme-linked immunosorbent assay, and the Ace-GST localization in adult worm was determined using the immunofluorescence method. The results showed that the full-length cDNA encoding Ace-GST was 468 bp, which had the highest homology with Ac-GST-1 (60.1%) and clustered into one branch (v-class) with Ac-GST-1 and Na-GST-1 in a phylogenetic tree. Mice immunized with recombinant Ace-GST showed specific IgG antibody response. Immunolocalization revealed that natural Ace-GST is mainly located in the epidermis, muscle and intestine of the adult. These results may lay a foundation for further studies on the biological function of Ace-GST.

Keywords

Ancylostoma ceylanicumglutathione S-transferasemolecular characterizationimmunolocalization
Type
Research Paper
Information
Journal of Helminthology , Volume 94 , 2020 , e118
Copyright
Copyright © Cambridge University Press 2020

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

Albonico, M, Bickle, Q, Ramsan, M, Montresor, A, Savioli, L and Taylor, M (2003) Efficacy of mebendazole and levamisole alone or in combination against intestinal nematode infections after repeated targeted mebendazole treatment in Zanzibar. Bulletin of the World Health Organization 81, 343–352.Google ScholarPubMed
Bowman, DD, Montgomery, SP, Zajac, AM, Eberhard, ML and Kazacos, KR (2010) Hookworms of dogs and cats as agents of cutaneous larva migrans. Trends in Parasitology 26, 162–167.CrossRefGoogle ScholarPubMed
Brophy, PM and Barrett, J (1990) Glutathione transferase in helminthes. Parasitology 2, 345–349.CrossRefGoogle Scholar
Brophy, PM and Pritchard, DI (1994) Parasitic helminth glutathione S-transferases: an update on their potential as targets for immuno- and chemotherapy. Experimental Parasitology 79, 89–96.CrossRefGoogle ScholarPubMed
Cervi, L, Rossi, G and Masih, DT (1999) Potential role for excretory-secretory forms of glutathione S-transferase (GST) in Fasciola hepatica. Parasitology 119, 627–633.CrossRefGoogle Scholar
Crompton, DWT (2000) The public health importance of hookworm disease. Parasitology 126, S39.CrossRefGoogle Scholar
Deponte, M and Becker, K (2005) Glutathione S-transferase from malarial parasites: structural and functional aspects. Methods in Enzymology 401, 241–253.CrossRefGoogle ScholarPubMed
Don, TA, Oksov, Y, Lustigman, S and Loukas, A (2007) Saposin-like proteins from the intestine of the blood-feeding hookworm, Ancylostoma caninum. Parasitology 134, 427–436.CrossRefGoogle ScholarPubMed
Hayes, RD, Flanagan, JU and Jowsey, IR (2005) Glutathione transferases. Annual Review of Pharmacology and Toxicology 45, 51–88.CrossRefGoogle ScholarPubMed
Hotez, PJ, Bethony, J, Bottazzi, ME and Buss, P (2005) Hookworm: the great infection of mankind. PLoS Medicine 2, e67.CrossRefGoogle ScholarPubMed
Jani, D, Nagarkatti, R, Beatty, W, Angel, R and Slebodnick, C (2008) HDP—a novel heme detoxification protein from the malaria parasite. PLoS Pathogens 4, e1000053.CrossRefGoogle ScholarPubMed
KampköTter, A, Volkmann, TE, Castro, SHD, Leiers, B, Klotz, L, Johnson, TE, Link, CD and Henkle-Dührsen, K (2003) Functional analysis of the glutathione S-transferase 3 from Onchocerca volvulus (Ov-GST-3): a parasite GST confers increased resistance to oxidative stress in Caenorhabditis elegans. Journal of Molecular Biology 325, 25–37.CrossRefGoogle ScholarPubMed
Klonis, N, Dilanian, R, Hanssen, E, Darmanin, C, Streltsov, V, Deed, S, Quiney, H and Tilley, L (2011) Hematin-hematin self-association states involved in the formation and reactivity of the malaria parasite pigment, hemozoin. Biochemistry 49, 6804–6811.CrossRefGoogle Scholar
Lathrop, JT and Timko, MP (1993) Regulation by heme of mitochondrial protein transport through a conserved amino acid motif. Science 259, 522–525.CrossRefGoogle ScholarPubMed
Liu, Y, Zheng, G, Alsarakibi, M, et al. (2013) Molecular identification of Ancylostoma caninum isolated from cats in southern China based on complete ITS sequence. BioMed Research International 2013, e86805.CrossRefGoogle ScholarPubMed
Murphy, JF (2013) The global burden of disease. Irish Medical Journal 106, 4.Google ScholarPubMed
Nakajima, O, Takahashi, S, Harigae, H, Furuyama, K, Hayashi, N, Sassa, S and Yamamoto, M (1999) Heme deficiency in erythroid lineage causes differentiation arrest and cytoplasmic iron overload. The European Molecular Biology Organization Journal 18, 6282–6289.CrossRefGoogle ScholarPubMed
Ngui, R, Lim, YAL, Traub, R, Mahmud, R and Mistam, MS (2012) Epidemiological and genetic data supporting the transmission of Ancylostoma ceylanicum among human and domestic animals. PLoS Neglected Tropical Diseases 6, e1522.CrossRefGoogle ScholarPubMed
Oakey, A (2011) Glutathione transferases: A structural perspective. Drug Metabolism Reviews 43, 138–151.CrossRefGoogle Scholar
Qi, Z, Hamza, I and O'Brian, MR (1999) Heme is an effector molecule for iron-dependent degradation of the bacterial iron response regulator (Irr) protein. Proceeding of National Academy of Science of the United States of America 96, 13056–13061.CrossRefGoogle ScholarPubMed
Skelly, PJ, Da'dara, AA, Li, XH, Castro-Borges, W and Wilson, RA (2014) Schistosome feeding and regurgitation. PLoS Pathogens 10, e1004246.CrossRefGoogle ScholarPubMed
Sun, J, Hoshino, H, Takaku, K, et al. (2002) Hemoprotein Bach1 regulates enhancer availability of heme oxygenase-1 gene. The European Molecular Biology Organization Journal 21, 5216–5224.CrossRefGoogle ScholarPubMed
Torres-Rivera, A and Landa, A (2008) Glutathione transferases from parasites: A biochemical view. Acta Tropica 105, 99–112.CrossRefGoogle ScholarPubMed
Traub, RJ, Inpankaew, T, Sutthikornchai, C, Sukthana, Y and Thompson, RCA (2008) PCR-based coprodiagnostic tools reveal dogs as reservoirs of zoonotic ancylostomiasis caused by Ancylostoma ceylanicum in temple communities in Bangkok. Veterinary Parasitology 155, 67–73.CrossRefGoogle ScholarPubMed
van Rossum, AJ, Jefferies, JR, Rijsewijk, FA, LaCourse, EJ, Teesdale-Spittle, P, Barrett, J, Tait, A and Brophy, PM (2004) Binding of hematin by a new class of glutathione transferase from the blood-feeding parasitic nematode Haemonchus contortus. Infection & Immunity 72, 2780–2790.CrossRefGoogle ScholarPubMed
Vega-Angeles, VT, Terrazas, LI, Ledesma-Soto, Y, Jimenez, L and Landa, A (2019) Taenia solium glutathione transferase fraction activates macrophages and favors the development of Th1-type response. Bioscience Reports 39, 20181132.CrossRefGoogle ScholarPubMed
Wei, JF, Damania, A, Gao, X, et al. (2016) The hookworm Ancylostoma ceylanicum intestinal transcriptome provides a platform for selecting drug and vaccine candidates. Parasites & Vectors 9, e518.CrossRefGoogle ScholarPubMed
Zhan, B, Liu, S, Perally, S, et al. (2005) Biochemical characterization and vaccine potential of a heme-binding glutathione transferase from the adult hookworm Ancylostoma caninum. Infection & Immunity 73, 6903–6911.CrossRefGoogle ScholarPubMed
Zhan, B, Perally, S, Brophy, PM, et al. (2010) Molecular cloning, biochemical characterization, and partial protective immunity of the heme-binding glutathione S-transferases from the human hookworm Necator americanus. Infection & Immunity 78, 1552–1563.CrossRefGoogle ScholarPubMed