Immunocytochemical evidence for the acidic nature of peroxisomes in methylotrophic yeasts

Hans R. Waterham, Ineke Keizer-Gunnink, Joel M. Goodman, Wim Harder, Marten Veenhuis

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The possible acidic nature of the peroxisomal matrix present in intact yeast cells was studied immunocytochemically, using the weak base DAMP as a probe. Spheroplasts of methanol-grown Candida boidinii and Hansenula polymorpha were regenerated and incubated with DAMP. After immunogold labelling, using antibodies against DAMP, a specific accumulation of gold particles was observed on the peroxisomal profiles. This labelling was absent in controls, performed in the presence of ionophores or chloroquine. These results support earlier observations, that in intact cells a pH-gradient exists across the peroxisomal membrane. Experiments, carried out on osmotically swollen spheroplasts indicated that maintenance of this pH-gradient is strongly related to the cell's integrity.

Original languageEnglish (US)
Pages (from-to)17-19
Number of pages3
JournalFEBS Letters
Volume262
Issue number1
DOIs
StatePublished - Mar 12 1990

Fingerprint

Peroxisomes
Yeast
Spheroplasts
Labeling
Proton-Motive Force
Yeasts
Candida
Ionophores
Chloroquine
Gold
Methanol
Pichia
Cells
Membranes
Antibodies
Maintenance
Experiments

Keywords

  • (Candida boidinii
  • Hansenula polymorpha)
  • Immunocytochemistry
  • Peroxisome
  • Proton gradient

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Immunocytochemical evidence for the acidic nature of peroxisomes in methylotrophic yeasts. / Waterham, Hans R.; Keizer-Gunnink, Ineke; Goodman, Joel M.; Harder, Wim; Veenhuis, Marten.

In: FEBS Letters, Vol. 262, No. 1, 12.03.1990, p. 17-19.

Research output: Contribution to journalArticle

Waterham, Hans R. ; Keizer-Gunnink, Ineke ; Goodman, Joel M. ; Harder, Wim ; Veenhuis, Marten. / Immunocytochemical evidence for the acidic nature of peroxisomes in methylotrophic yeasts. In: FEBS Letters. 1990 ; Vol. 262, No. 1. pp. 17-19.
@article{ee1cabec61b7409eb20f71c40845e63f,
title = "Immunocytochemical evidence for the acidic nature of peroxisomes in methylotrophic yeasts",
abstract = "The possible acidic nature of the peroxisomal matrix present in intact yeast cells was studied immunocytochemically, using the weak base DAMP as a probe. Spheroplasts of methanol-grown Candida boidinii and Hansenula polymorpha were regenerated and incubated with DAMP. After immunogold labelling, using antibodies against DAMP, a specific accumulation of gold particles was observed on the peroxisomal profiles. This labelling was absent in controls, performed in the presence of ionophores or chloroquine. These results support earlier observations, that in intact cells a pH-gradient exists across the peroxisomal membrane. Experiments, carried out on osmotically swollen spheroplasts indicated that maintenance of this pH-gradient is strongly related to the cell's integrity.",
keywords = "(Candida boidinii, Hansenula polymorpha), Immunocytochemistry, Peroxisome, Proton gradient",
author = "Waterham, {Hans R.} and Ineke Keizer-Gunnink and Goodman, {Joel M.} and Wim Harder and Marten Veenhuis",
year = "1990",
month = "3",
day = "12",
doi = "10.1016/0014-5793(90)80142-6",
language = "English (US)",
volume = "262",
pages = "17--19",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Immunocytochemical evidence for the acidic nature of peroxisomes in methylotrophic yeasts

AU - Waterham, Hans R.

AU - Keizer-Gunnink, Ineke

AU - Goodman, Joel M.

AU - Harder, Wim

AU - Veenhuis, Marten

PY - 1990/3/12

Y1 - 1990/3/12

N2 - The possible acidic nature of the peroxisomal matrix present in intact yeast cells was studied immunocytochemically, using the weak base DAMP as a probe. Spheroplasts of methanol-grown Candida boidinii and Hansenula polymorpha were regenerated and incubated with DAMP. After immunogold labelling, using antibodies against DAMP, a specific accumulation of gold particles was observed on the peroxisomal profiles. This labelling was absent in controls, performed in the presence of ionophores or chloroquine. These results support earlier observations, that in intact cells a pH-gradient exists across the peroxisomal membrane. Experiments, carried out on osmotically swollen spheroplasts indicated that maintenance of this pH-gradient is strongly related to the cell's integrity.

AB - The possible acidic nature of the peroxisomal matrix present in intact yeast cells was studied immunocytochemically, using the weak base DAMP as a probe. Spheroplasts of methanol-grown Candida boidinii and Hansenula polymorpha were regenerated and incubated with DAMP. After immunogold labelling, using antibodies against DAMP, a specific accumulation of gold particles was observed on the peroxisomal profiles. This labelling was absent in controls, performed in the presence of ionophores or chloroquine. These results support earlier observations, that in intact cells a pH-gradient exists across the peroxisomal membrane. Experiments, carried out on osmotically swollen spheroplasts indicated that maintenance of this pH-gradient is strongly related to the cell's integrity.

KW - (Candida boidinii

KW - Hansenula polymorpha)

KW - Immunocytochemistry

KW - Peroxisome

KW - Proton gradient

UR - http://www.scopus.com/inward/record.url?scp=0025212105&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025212105&partnerID=8YFLogxK

U2 - 10.1016/0014-5793(90)80142-6

DO - 10.1016/0014-5793(90)80142-6

M3 - Article

C2 - 2156731

AN - SCOPUS:0025212105

VL - 262

SP - 17

EP - 19

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1

ER -