Abstract
Monoclonal antibodies were raised against pathogenic promastigotes ofLeishmania donovani of Indian origin. Among these, one was used for immuno-affinity purification of a 78 kDa membrane protein present in both the amastigote and promastigote forms of the parasite. Results of immunoblot experiments with the anti-78 kDa antibody revealed that the protein was present only in parasites belonging to theL. donovani complex. The expression of the protein was observed to be the same during different phases of growth of the promastigotes. Therefore, the 78 kDa protein is neither stage-specific nor differentially regulated. Surface iodination and subcellular fractionation of the promastigotes indicated that the protein was localized on the cell surface. The 78 kDa protein was found to inhibit the binding of promastigotes to macrophages significantly, suggesting that it may play a role in the process of infection. Thus, here we report the purification of a surface protein ofL. donovani of Indian origin, which may play an important role in the process of infection.
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Abbreviations
- BCIP:
-
5-bromo-4-chloro-3-indolyl phosphate
- BSA:
-
bovine serum albumin
- DOC:
-
deoxycholic acid
- EGTA:
-
ethyleneglycol-bis-(beta-amino ethyl ether) N,N,N,N-tetra acetic acid
- EDTA:
-
ethylenediaminetetraacetate
- FACS:
-
fluorescence activated cell sorter
- HRP:
-
horse radish peroxidase
- KA:
-
kala-azar
- NBT:
-
nitrobluetetrazolium
- PBS:
-
phosphate buffer saline
- PMSF:
-
phenyl methyl sulfonyl fluoride
- PKDL:
-
post kala-azar dermal leishmaniasis
- SDS-PAGE:
-
sodium dodecyl sulphate polyacrylamide gel electrophoresis
- VL:
-
visceral leishmaniasis
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Mukherjee, M., Bhattacharyya, A. & Duttagupta, S. Monoclonal antibody affinity purification of a 78 kDa membrane protein ofLeishmania donovani of Indian origin and its role in host-parasite interaction. J Biosci 27, 665–672 (2002). https://doi.org/10.1007/BF02708374
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DOI: https://doi.org/10.1007/BF02708374