Differential targeting of two glucose transporters from Leishmania enriettii is mediated by an NH2-terminal domain

Robert C. Piper, Xiaowen Xu, David G. Russell, Brian M. Little, Scott Landfear

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Leishmania are parasitic protozoa with two major stages in their life cycle: flagellated promastigotes that live in the gut of the insect vector and non-flagellated amastigotes that live inside the lysosomes of the vertebrate host macrophages. The Pro-1 glucose transporter of L. enriettii exists as two isoforms, iso-1 and iso-2, which are both expressed primarily in the promastigote stage of the life cycle. These two iso-forms constitute modular structures: they differ exclusively and extensively in their NH2- terminal hydrophilic domains, but the remainder of each isoform sequence is identical to that of the other. We have localized these glucose transporters within promastigotes by two approaches. In the first method, we have raised a polyclonal antibody against the COOH-terminal hydrophilic domain shared by both iso-1 and iso-2, and we have used this antibody to detect the transporters by confocal immunofluorescence microscopy and immunoelectron microscopy. The staining observed with this antibody occurs primarily on the plasma membrane and the membrane of the flagellar pocket, but there is also light staining on the flagellum. We have also localized each isoform separately by introducing an epitope tag into each protein sequence. These experiments demonstrate that iso-1, the minor isoform, resides primarily on the flagellar membrane, while iso-2, the major isoform, is located on the plasma membrane and the flagellar pocket. Hence, each isoform is differentially sorted, and the structural information for targeting each transporter isoform to its correct membrane address resides within the NH2- terminal hydrophilic domain.

Original languageEnglish (US)
Pages (from-to)499-508
Number of pages10
JournalJournal of Cell Biology
Volume128
Issue number4
DOIs
StatePublished - Feb 1995

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Leishmania enriettii
Facilitative Glucose Transport Proteins
Protein Isoforms
Life Cycle Stages
Membranes
Antibodies
Cell Membrane
Insect Vectors
Staining and Labeling
Immunoelectron Microscopy
Flagella
Leishmania
Lysosomes
Fluorescence Microscopy
Confocal Microscopy
Vertebrates
Epitopes
Macrophages
Light

ASJC Scopus subject areas

  • Cell Biology

Cite this

Differential targeting of two glucose transporters from Leishmania enriettii is mediated by an NH2-terminal domain. / Piper, Robert C.; Xu, Xiaowen; Russell, David G.; Little, Brian M.; Landfear, Scott.

In: Journal of Cell Biology, Vol. 128, No. 4, 02.1995, p. 499-508.

Research output: Contribution to journalArticle

Piper, Robert C. ; Xu, Xiaowen ; Russell, David G. ; Little, Brian M. ; Landfear, Scott. / Differential targeting of two glucose transporters from Leishmania enriettii is mediated by an NH2-terminal domain. In: Journal of Cell Biology. 1995 ; Vol. 128, No. 4. pp. 499-508.
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