Molecular dissection of the functional domains of a unique, tartrate-resistant, surface membrane acid phosphatase in the primitive human pathogen Leishmania donovani.
The primitive trypanosomatid pathogen of humans, Leishmania donovani, constitutively expresses a unique externally oriented, tartrate-resistant, acid phosphatase on its surface membrane. This is of interest because these organisms are obligate intracellular protozoan parasites that reside and multiply within the hydrolytic milieu of mammalian macrophage phago-lysosomes. Here we report the identification of the gene encoding this novel L. donovani enzyme. In addition, we characterized its structure, demonstrated its constitutive expression in both parasite developmental forms, and determined the cell surface membrane localization of its translated protein product. Further, we used a variety of green fluorescent protein chimeric constructs as reporters in a homologous leishmanial expression system to dissect the functional domains of this unique, tartrate-resistant, surface membrane enzyme.
Shakarian
Alison M
AM
Cell Biology Section, Laboratory of Parasitic Diseases, NIAID, National Institutes of Health, Bethesda, Maryland 20892-0425, USA.
Joshi
Manju B
MB
Ghedin
Elodie
E
Dwyer
Dennis M
DM
D000135:Acid Phosphatase / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D007893:Leishmania donovani / Q000201:enzymology*; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013644:Tartrates / Q000494:pharmacology*
D016054:DNA, Protozoan; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D013644:Tartrates; D049452:Green Fluorescent Proteins; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D000135:Acid Phosphatase; C029768:tartaric acid