Advanced

A fluorescent probe of polyamine transport accumulates into intracellular acidic vesicles via a two-step mechanism

Soulet, Denis LU ; Gagnon, Bruno; Rivest, Serge; Audette, Marie and Poulin, Richard (2004) In Journal of Biological Chemistry 279(47). p.49355-49366
Abstract
Mammalian polyamine carriers have not yet been molecularly identified. The fluoroprobe Spd-C2-BODIPY faithfully reports polyamine transport and accumulates almost exclusively in polyamine-sequestering vesicles (PSVs). Polyamines might thus be imported first by a plasma membrane carrier and then sequestered into pre-existing PSVs (model A), or be directly captured by polyamine receptors undergoing endocytosis (model B). Spd-C2-BODIPY uptake was unaffected in receptor-mediated endocytosis-deficient Chinese hamster ovary cell mutants. PSVs strongly colocalized with acidic vesicles of the late endocytic compartment and the trans Golgi. Virtually perfect colocalization between PSVs and acidic vesicles was found in Chinese hamster ovary cell... (More)
Mammalian polyamine carriers have not yet been molecularly identified. The fluoroprobe Spd-C2-BODIPY faithfully reports polyamine transport and accumulates almost exclusively in polyamine-sequestering vesicles (PSVs). Polyamines might thus be imported first by a plasma membrane carrier and then sequestered into pre-existing PSVs (model A), or be directly captured by polyamine receptors undergoing endocytosis (model B). Spd-C2-BODIPY uptake was unaffected in receptor-mediated endocytosis-deficient Chinese hamster ovary cell mutants. PSVs strongly colocalized with acidic vesicles of the late endocytic compartment and the trans Golgi. Virtually perfect colocalization between PSVs and acidic vesicles was found in Chinese hamster ovary cell mutants that are blocked either in the late endosome/lysosome fusion process or in the maturation of multivesicular bodies. Prior inhibition of the V-ATPase dramatically decreased total Spd-C2-BODIPY accumulation while increasing cytosolic fluorescence. Conversely, cells pre-loaded with the probe slowly released it from PSVs upon V-ATPase inhibition. The present data thus support model A, and indicate that polyamine accumulation is primarily driven by the activity of a vesicular H+:polyamine carrier. (Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
279
issue
47
pages
49355 - 49366
publisher
ASBMB
external identifiers
  • pmid:15208319
  • scopus:10444227401
ISSN
1083-351X
DOI
10.1074/jbc.M401287200
language
English
LU publication?
no
id
55243eab-c278-412a-9e6b-941d1648b9d7 (old id 1130921)
date added to LUP
2008-06-18 13:30:52
date last changed
2017-11-12 03:28:28
@article{55243eab-c278-412a-9e6b-941d1648b9d7,
  abstract     = {Mammalian polyamine carriers have not yet been molecularly identified. The fluoroprobe Spd-C2-BODIPY faithfully reports polyamine transport and accumulates almost exclusively in polyamine-sequestering vesicles (PSVs). Polyamines might thus be imported first by a plasma membrane carrier and then sequestered into pre-existing PSVs (model A), or be directly captured by polyamine receptors undergoing endocytosis (model B). Spd-C2-BODIPY uptake was unaffected in receptor-mediated endocytosis-deficient Chinese hamster ovary cell mutants. PSVs strongly colocalized with acidic vesicles of the late endocytic compartment and the trans Golgi. Virtually perfect colocalization between PSVs and acidic vesicles was found in Chinese hamster ovary cell mutants that are blocked either in the late endosome/lysosome fusion process or in the maturation of multivesicular bodies. Prior inhibition of the V-ATPase dramatically decreased total Spd-C2-BODIPY accumulation while increasing cytosolic fluorescence. Conversely, cells pre-loaded with the probe slowly released it from PSVs upon V-ATPase inhibition. The present data thus support model A, and indicate that polyamine accumulation is primarily driven by the activity of a vesicular H+:polyamine carrier.},
  author       = {Soulet, Denis and Gagnon, Bruno and Rivest, Serge and Audette, Marie and Poulin, Richard},
  issn         = {1083-351X},
  language     = {eng},
  number       = {47},
  pages        = {49355--49366},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {A fluorescent probe of polyamine transport accumulates into intracellular acidic vesicles via a two-step mechanism},
  url          = {http://dx.doi.org/10.1074/jbc.M401287200},
  volume       = {279},
  year         = {2004},
}