The major surface protease (MSP or GP63) in the intracellular amastigote stage of Leishmania chagasi

Chia-Hung Christine Hsiao; Chaoqun Yao; Patricia Storlie; John E Donelson; Mary E Wilson

doi:10.1016/j.molbiopara.2007.10.008

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The major surface protease (MSP or GP63) in the intracellular amastigote stage of Leishmania chagasi

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The major surface protease (MSP or GP63) in the intracellular amastigote stage of Leishmania chagasi

Chia-Hung Christine Hsiao, Chaoqun Yao, Patricia Storlie, John E Donelson and Mary E Wilson

Molecular and biochemical parasitology, Vol.157(2), pp.148-159

2008

DOI: 10.1016/j.molbiopara.2007.10.008

PMCID: PMC2713036

PMID: 18067978

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Abstract

The Leishmania spp. protozoa have an abundant surface metalloprotease called MSP (major surface protease), which in Leishmania chagasi is encoded by three distinct gene classes ( MSPS, MSPL, MSPC). Although MSP has been characterized primarily in extracellular promastigotes, it also facilitates survival of intracellular amastigotes. Promastigotes express MSPS, MSPL, and two forms of MSPC RNAs, whereas amastigotes express only MSPL RNA and one MSPC transcript. We confirmed the presence of MSPC protein in both promastigotes and amastigotes by liquid chromatography–tandem mass spectrometry (LC–MS/MS). More than 10 MSP isoforms were visualized in both amastigotes and promastigotes using two-dimensional immunoblots, but amastigote MSPs migrated at a more acidic p I. Promastigote MSPs were N-glycosylated, whereas most amastigote MSPs were not. Immuno-electron microscopy showed that two-thirds of the promastigote MSP is distributed along the cell surface. In contrast, most amastigote MSP localized at the flagellar pocket, the major site of leishmania endocytosis/exocytosis. Biochemical analyses indicated that most amastigote MSP is soluble in the cytosol, vesicles or organelles, whereas most promastigote MSP is membrane-associated and GPI anchored. Activity gels and immunoblots confirmed the presence of a novel proteolytically active amastigote MSP of higher Mr than the promastigote MSPs. Furthermore, promastigote MSP is shed extracellularly whereas MSP is not shed from axenic amastigotes. We conclude that amastigotes and promastigotes both express multiple MSP isoforms, but these MSPs differ biochemically and localize differently in the two parasite stages. We hypothesize that MSP plays different roles in the extracellular versus intracellular forms of Leishmania spp.

Metalloprotease

Flagellar pocket

Protease

Amastigote

Leishmania

Details

Title: Subtitle: The major surface protease (MSP or GP63) in the intracellular amastigote stage of Leishmania chagasi
Creators: Chia-Hung Christine Hsiao - Molecular Biology Program, University of Iowa, Iowa City, IA, United States
Chaoqun Yao - Department of Internal Medicine, University of Iowa, Iowa City, IA, United States
Patricia Storlie - Microbiology, University of Iowa, Iowa City, IA, United States
John E Donelson - Molecular Biology Program, University of Iowa, Iowa City, IA, United States
Mary E Wilson - Molecular Biology Program, University of Iowa, Iowa City, IA, United States
Resource Type: Journal article
Publication Details: Molecular and biochemical parasitology, Vol.157(2), pp.148-159
DOI: 10.1016/j.molbiopara.2007.10.008
PMID: 18067978
PMCID: PMC2713036
NLM abbreviation: Mol Biochem Parasitol
ISSN: 0166-6851
eISSN: 1872-9428
Publisher: Elsevier B.V
Language: English
Date published: 2008
Academic Unit: Microbiology and Immunology; International Programs; Epidemiology; Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9984001133302771

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