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Structurally conserved soluble acid phosphatases are synthesized and released by Leishmania major promastigotes
1Cell Biology Section, Laboratory of Parasitic Disease, Division of Intramural Research, The National Institute of Allergy and Infectious Diseases, The National Institutes of Health, Building 4, Room 126, 4 Center Drive MSC 0425, Bethesda, MD, USA.
Abstract:
Previously it was reported that promastigotes of virtually all pathogenic Leishmania species, except Leishmania major, release a structurally conserved soluble acid phosphatase (AcP) activity during their growth in vitro (P. S. Doyle and D. M. Dwyer, Exp. Parasitol. 77, 435-444 1993). In the current study we used a highly sensitive fluorogenic detection method to demonstrate that soluble AcPs were in fact produced by promastigotes of several different strains of L. major. These L. major AcP activities were readily immunoprecipitated with a rabbit antibody previously generated against the L. donovani AcP. Results of metabolic labeling and immunoprecipitations demonstrated that AcPs produced by the L. majors strains examined had an apparent molecular mass of approximately 77 kDa. Results of Southern hybridization analysis with an L. donovani AcP gene probe showed that the AcP gene loci were conserved in the L. major strains examined. Taken together, these results indicate that the AcP enzyme has been structurally and functionally conserved throughout the evolution of pathogenic species of Leishmania. Such conservation suggests that the AcPs play a functional role in the growth and survival of this group of important human pathogens.
Insights
Pathogenic Leishmania parasites, including Leishmania major, produce a conserved soluble acid phosphatase (AcP) enzyme. This enzyme is crucial for the growth and survival of these important human pathogens.
Area of Science:
- Parasitology
- Molecular Biology
- Biochemistry
Background:
- Soluble acid phosphatase (AcP) activity was previously reported in most pathogenic Leishmania species.
- Leishmania major was an exception, with no reported AcP activity during in vitro growth.
Purpose of the Study:
- To investigate the production of soluble AcPs by Leishmania major.
- To determine the structural and functional conservation of AcP in pathogenic Leishmania species.
Main Methods:
- Utilized a sensitive fluorogenic detection method to identify AcP activity.
- Employed immunoprecipitation with anti-L. donovani AcP antibody.
- Conducted metabolic labeling and Southern hybridization analysis using an L. donovani AcP gene probe.
Main Results:
- Demonstrated soluble AcP production by promastigotes of multiple L. major strains.
- Confirmed AcP activity in L. major is immunoprecipitable with anti-L. donovani AcP antibody.
- Identified an approximate 77 kDa molecular mass for L. major AcPs and conserved AcP gene loci.
Conclusions:
- Soluble acid phosphatase (AcP) is produced by Leishmania major.
- The AcP enzyme is structurally and functionally conserved across pathogenic Leishmania species.
- Conserved AcPs likely play a vital role in the growth and survival of Leishmania pathogens.