Ecotin-like serine peptidase inhibitor ISP1 of Leishmania major plays a role in flagellar pocket dynamics and promastigote differentiation

Lesley S Morrison, Amy Goundry, Marilia S Faria, Laurence Tetley, Sylvain C Eschenlauer, Gareth D Westrop, Anna Dostalova, Petr Volf, Graham H Coombs, Ana Paula C A Lima, Jeremy C Mottram

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Leishmania ISPs are ecotin-like natural peptide inhibitors of trypsin-family serine peptidases, enzymes that are absent from the Leishmania genome. This led to the proposal that ISPs inhibit host serine peptidases and we have recently shown that ISP2 inhibits neutrophil elastase, thereby enhancing parasite survival in murine macrophages. In this study we show that ISP1 has less serine peptidase inhibitory activity than ISP2, and in promastigotes both are generally located in the cytosol and along the flagellum. However, in haptomonad promastigotes there is a prominent accumulation of ISP1 and ISP2 in the hemidesmosome and for ISP2 on the cell surface. An L. major mutant deficient in all three ISP genes (Δisp1/2/3) was generated and compared with Δisp2/3 mutants to elucidate the physiological role of ISP1. In in vitro cultures, the Δisp1/2/3 mutant contained more haptomonad, nectomonad and leptomonad promastigotes with elongated flagella and reduced motility compared with Δisp2/3 populations, moreover it was characterized by very high levels of release of exosome-like vesicles from the flagellar pocket. These data suggest that ISP1 has a primary role in flagellar homeostasis, disruption of which affects differentiation and flagellar pocket dynamics.
LanguageEnglish
JournalCellular Microbiology
Early online date8 May 2012
DOIs
Publication statusE-pub ahead of print - 8 May 2012

Fingerprint

Leishmania major
Protease Inhibitors
Serine
Peptide Hydrolases
Flagella
Leishmania
Hemidesmosomes
Exosomes
Leukocyte Elastase
Trypsin Inhibitors
Cytosol
Parasites
Homeostasis
Macrophages
Genome
Peptides
Enzymes
Population
Genes

Keywords

  • Leishmania ISP
  • Leishmania genome
  • serine peptidases

Cite this

Morrison, Lesley S ; Goundry, Amy ; Faria, Marilia S ; Tetley, Laurence ; Eschenlauer, Sylvain C ; Westrop, Gareth D ; Dostalova, Anna ; Volf, Petr ; Coombs, Graham H ; Lima, Ana Paula C A ; Mottram, Jeremy C. / Ecotin-like serine peptidase inhibitor ISP1 of Leishmania major plays a role in flagellar pocket dynamics and promastigote differentiation. In: Cellular Microbiology. 2012.
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Ecotin-like serine peptidase inhibitor ISP1 of Leishmania major plays a role in flagellar pocket dynamics and promastigote differentiation. / Morrison, Lesley S; Goundry, Amy; Faria, Marilia S; Tetley, Laurence; Eschenlauer, Sylvain C; Westrop, Gareth D; Dostalova, Anna; Volf, Petr; Coombs, Graham H; Lima, Ana Paula C A; Mottram, Jeremy C.

In: Cellular Microbiology, 08.05.2012.

Research output: Contribution to journalArticle

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