Active transport in L. donovani is dependent on a proton gradient across the plasma membrane that is established through the action of an ATPase that extrudes protons. This proton must also play a vital role in the survival of the parasite within the acidic environment of the vertebrate host's phagolysosomes. Our long term goal is to understand the structure, function and regulation of this enzyme and, through this understanding to provide avenues to the control of this important pathogen.
The aim of this proposal is to apply molecular cloning technology to produce molecular probes with which to analyze the macromolecules that mediate the proton pumping system. We have succeeded in the first phase of this endeavor and have cloned parts of two tandemly arrayed genes that are structurally capable of encoding plasma membrane cation pumps. We will clone this tandem pair of L. donovani genes in their entirety. We will determine the DNA sequences of the genes, and predict the amino acid sequences of their protein products. This sequence information will be used produce nucleic acid and antibody probes for the analysis of expression in the two life cycle stages of the parasite. We will compare the abundance of these gene products to proton pumping activity, and examine the enzymatic activities of the proteins encoded by the presumptive proton pump genes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM039622-02
Application #
3296771
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1988-02-01
Project End
1993-01-31
Budget Start
1989-02-01
Budget End
1990-01-31
Support Year
2
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Jiang, Suping; Meadows, Juliana; Anderson, Steven A et al. (2002) Antileishmanial activity of the antiulcer agent omeprazole. Antimicrob Agents Chemother 46:2569-74
Jiang, S; Anderson, S A; Winget, G D et al. (1994) Plasma membrane K+/H(+)-ATPase from Leishmania donovani. J Cell Physiol 159:60-6
Anderson, S A; Mukkada, A J (1994) Biochemical and immunochemical characterization of a P-type ATPase from Leishmania donovani promastigote plasma membrane. Biochim Biophys Acta 1195:71-80
Anderson, S A; Jiang, S; Mukkada, A J (1994) The beta-aspartyl phosphate intermediate in a Leishmania donovani promastigote plasma membrane P-type ATPase. Biochim Biophys Acta 1195:81-8
Liang, K Y; Zeger, S L (1993) Regression analysis for correlated data. Annu Rev Public Health 14:43-68
Glaser, T A; Mukkada, A J (1992) Proline transport in Leishmania donovani amastigotes: dependence on pH gradients and membrane potential. Mol Biochem Parasitol 51:1-8
Glaser, T A; Utz, G L; Mukkada, A J (1992) The plasma membrane electrical gradient (membrane potential) in Leishmania donovani promastigotes and amastigotes. Mol Biochem Parasitol 51:9-15
Meade, J C; Coombs, G H; Mottram, J C et al. (1991) Conservation of cation-transporting ATPase genes in Leishmania. Mol Biochem Parasitol 45:29-38
Meade, J C; Hudson, K M; Stringer, S L et al. (1989) A tandem pair of Leishmania donovani cation transporting ATPase genes encode isoforms that are differentially expressed. Mol Biochem Parasitol 33:81-91
Glaser, T A; Baatz, J E; Kreishman, G P et al. (1988) pH homeostasis in Leishmania donovani amastigotes and promastigotes. Proc Natl Acad Sci U S A 85:7602-6