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Proteomic examination of Leishmania chagasi plasma membrane proteins: contrast between avirulent and virulent (metacyclic) parasite forms
Journal article   Open access   Peer reviewed

Proteomic examination of Leishmania chagasi plasma membrane proteins: contrast between avirulent and virulent (metacyclic) parasite forms

Chaoqun Yao, Yalan Li, John E Donelson and Mary E Wilson
Proteomics. Clinical applications, Vol.4(1), pp.4-16
01/2010
DOI: 10.1002/prca.200900050
PMCID: PMC3049999
PMID: 21137013
url
https://www.ncbi.nlm.nih.gov/pmc/articles/3049999View
Open Access

Abstract

Leishmaniasis annually leads to two million new cases with 59 thousand deaths worldwide. Promastigotes of the causative Leishmania spp. develop from procyclic (log) to the highly virulent metacyclic stage within the sand fly vector. We hypothesized that proteins important for promastigote virulence might be uniquely represented in the plasma membrane of metacyclic, but not log, promastigotes. Purified metacyclic promastigotes from stationary phase cultures of Leishmania chagasi were applied to prepare membrane preparations either by surface biotinylation-streptavidin affinity separation or by octyl glucoside detergent extraction. These membrane fractions were enriched over 130 and 250 fold, respectively, estimated by western blotting for the plasma membrane marker MSP. A total of 447 or 33 proteins were identified by surface biotinylation or detergent extraction, respectively, by LC-MS/MS. Confocal microscopy suggested the difference between the lists was due to the fact that proteins localized both on the surface membrane and within the flagellar pocket were accessible to surface biotinylation. Using detergent extraction, we found different proteins were present in membrane proteins of logarithmic stage compared to metacyclic stage promastigotes. Several dozens were stage specific. These data provide a foundation for identifying virulence factors in the plasma membranes of Leishmania spp. promastigotes during metacyclogenesis.
plasma membrane proteins proteomics metacyclic promastigotes Leishmania

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