Multiple products of the Leishmania chagasi major surface protease (MSP or GP63) gene family

Yao, Chaoqun; Luo, Jiwen; Storlie, Patricia; Donelson, John E.; Wilson, Mary E.

doi:10.1016/j.molbiopara.2004.03.010

Cited by 21 publications

(41 citation statements)

References 49 publications

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“…Although RFLP analysis seems to be successful for species identification, the obtained patterns can be complicated and difficult to interpret because of so-called isogenes. These are variations between copies within the same parasite (169,175), which also render sequencing of the gp63 target impractical. Also, because antigen genes are under constant pressure from the immune system (176, 177), they are primarily suited for exploring clinical pleomorphism at the intraspecies level rather than for typing species.…”

Section: Gp63mentioning

confidence: 99%

Species Typing in Dermal Leishmaniasis

2015

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SUMMARY Leishmania is an infectious protozoan parasite related to African and American trypanosomes. All Leishmania species that are pathogenic to humans can cause dermal disease. When one is confronted with cutaneous leishmaniasis, identification of the causative species is relevant in both clinical and epidemiological studies, case management, and control. This review gives an overview of the currently existing and most used assays for species discrimination, with a critical appraisal of the limitations of each technique. The consensus taxonomy for the genus is outlined, including debatable species designations. Finally, a numerical literature analysis is presented that describes which methods are most used in various countries and regions in the world, and for which purposes.

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Section: Gp63mentioning

confidence: 99%

Species Typing in Dermal Leishmaniasis

2015

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“…The attenuated L5 strain of L. chagasi has been continuously cultured in vitro in HOMEM for over 9 years (43). Strain L5 differs from the virulent strain in several respects, including (i) lessabundant MSP and the expression of only MSPL genes (6,43,48), (ii) a shorter and simpler lipophosphoglycan (27), and (iii) reduced virulence for rodent models (43). In some experiments, virulent promastigotes were spread on semisolid M199 agar plates to obtain clonal cells (19).…”

Section: Parasitesmentioning

confidence: 99%

“…Both cells and cell-free spent medium were collected. Newly synthesized MSP, localized either on the cell surface or intracellularly, was isolated by streptavidin-affinity purification and immunoprecipitated from the streptavidin-cleared fraction, respectively, and detected by autoradiography as previously described (47,48). The efficiency of pull-down via biotinstreptavidin was routinely monitored by peroxidase-conjugated ExtrAvidin (Sigma) and ECL Western blotting detection reagents (Amersham).…”

Section: Parasitesmentioning

confidence: 99%

“…To investigate whether membrane lipid chelation reagent M␤CD affects the release of surface-localized MSP, stationary-phase promastigotes were incubated in either 0 or 15 mM M␤CD for 3 h after surface biotinylation. Spent medium was collected and concentrated as previously described (48). Both biotinylated proteins and the internal MSP were analyzed.…”

Section: Parasitesmentioning

confidence: 99%

“…During in vitro promastigote growth of virulent strain L. chagasi from the logarithmic to the stationary phase, MSP protein abundance increases 14-fold. Concomitantly, the number of MSP isoforms observed on two-dimensional gel electrophoresis (2-DE) increases from 4 to 11 (46)(47)(48). In the present study we use "MSP" or "MSPs" when referring to properties of all MSP isoforms and "MSP isoforms" when referring to specific MSP isoforms.…”

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Internal and Surface-Localized Major Surface Proteases ofLeishmaniaspp. and Their Differential Release from Promastigotes

2007

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Major surface protease (MSP), also called GP63, is a virulence factor of Leishmania spp. protozoa. There are three pools of MSP, located either internally within the parasite, anchored to the surface membrane, or released into the extracellular environment. The regulation and biological functions of these MSP pools are unknown. We investigated here the trafficking and extrusion of surface versus internal MSPs. Virulent Leishmania chagasi undergo a growth-associated lengthening in the t 1/2 of surface-localized MSP, but this did not occur in the attenuated L5 strain. The release of surface-localized MSP was enhanced in a dose-dependent manner by M␤CD, which chelates membrane cholesterol-ergosterol. Furthermore, incubation of promastigotes at 37°C with Matrigel matrix, a soluble basement membrane extract of Engelbreth-Holm-Swarm tumor cells, stimulated the release of internal MSP but not of surface-located MSP. Taken together, these data indicate that MSP subpopulations in distinct cellular locations are released from the parasite under different environmental conditions. We hypothesize that the internal MSP with its lengthy t 1/2 does not serve as a pool for promastigote surface MSP in the sand fly vector but that it instead functions as an MSP pool ready for quick release upon inoculation of metacyclic promastigotes into mammals. We present a model in which these different MSP pools are released under distinct life cycle-specific conditions.The digenetic protozoa of Leishmania spp. shuttle between an extracellular promastigote form in the sand fly vector and an intracellular amastigote form in mammalian hosts, including humans. In the sand fly, the avirulent procyclic promastigotes develop to the virulent metacyclic organisms, a process termed metacyclogenesis that can be mimicked by in vitro cultivation of logarithmic-to stationary-phase promastigotes (36). Leishmania causes 1.5 to 2 million new cases of human leishmaniasis annually, with manifestations ranging from selfhealing cutaneous sores to life-threatening visceral leishmaniasis (8, 9). Among a few well-characterized Leishmania virulence factors is the major surface protease (MSP), also called GP63. MSP plays several important roles during Leishmania spp. infection of mammals, including (i) enhancing promastigote phagocytosis by macrophages, (ii) facilitating promastigote evasion of complement-mediated lysis, and (iii) promoting amastigote survival in the phagolysosomes of macrophages (see reference 45 for a review). There is also evidence suggesting that in the sand fly MSP plays a role in the early-stage development of promastigotes (16) and contributes to promastigote adhesion in the guts and salivary glands (see reference 37 for a review).MSP is encoded by a family of highly conserved genes organized in a tandem array. MSP genes (MSPs) and homologues have been found in all Leishmania spp. studied to date, as well as in other trypanosomatids, including the monoxenous insect parasite Crithidia and the extracellular mammalian parasite Trypanosoma brucei...

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

Yao

Donelson

et al. 2010

Proteomics Clinical Apps

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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.

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Multiple products of the Leishmania chagasi major surface protease (MSP or GP63) gene family

Cited by 21 publications

References 49 publications

Species Typing in Dermal Leishmaniasis

Species Typing in Dermal Leishmaniasis

Internal and Surface-Localized Major Surface Proteases ofLeishmaniaspp. and Their Differential Release from Promastigotes

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

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