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Permeability properties of peroxisomal membranes from yeasts

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Abstract

We have studied the permeability properties of intact peroxisomes and purified peroxisomal membranes from two methylotrophic yeasts. After incorporation of sucrose and dextran in proteoliposomes composed of asolectin and peroxisomal membranes isolated from the yeasts Hansenula polymorpha and Candida boidinii a selective leakage of sucrose occurred indicating that the peroxisomal membranes were permeable to small molecules. Since the permeability of yeast peroxisomal membranes in vitro may be due to the isolation procedure employed, the osmotic stability of peroxisomes was tested during incubations of intact protoplasts in hypotonic media. Mild osmotic swelling of the protoplasts also resulted in swelling of the peroxisomes present in these cells but not in a release of their matrix proteins. The latter was only observed when the integrity of the cells was disturbed due to disruption of the cell membrane during further lowering of the concentration of the osmotic stabilizer. Stability tests with purified peroxisomes indicated that this leak of matrix proteins was not associated with the permeability to sucrose. Various attempts to mimic the in vivo situation and generate a proton motive force across the peroxisomal membranes in order to influence the permeability properties failed. Two different proton pumps were used for this purpose namely bacteriorhodopsin (BR) and reaction center-light-harvesting complex I (RCLHI complex). After introduction of BR into the membrane of intact peroxisomes generation of a pH-gradient was not or barely detectable. Since this pump readily generated a pH-gradient in pure liposomes, these results strengthened the initial observations on the leakiness of the peroxisomal membrane fragments. Generation of a membrane potential (Δψ) was also not observed when RCLHI complex was introduced into vesicles of purified peroxisomal membranes. The significance of the observed permeability of isolated yeast peroxisomal membranes to small molecules with respect to current and future in vitro import studies is discussed.

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Abbreviations

CL:

cardiolinin

PE:

phosphatidylethanolamine

PC:

phosphatidylcholine

MES:

2-(N-Morpholino)ethanesulfonic acid

R18 :

octadecyl Rhodamine B Chloride

SUVs:

small unilamellar vesicles

RCLHI-complex:

reaction center-light-harvesting complex I

BR:

bacteriorhodopsin

DCCD:

N,N′-dicyclohexylcarbodiimide

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Author notes

  1. A. C. Douma

    Present address: Center for Phytotechnology, RUL/TNO, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands

Authors and Affiliations

  1. Department of Microbiology, Biological Centre, University of Groningen, Kerklaan 30, NL-9751 NN, Haren, The Netherlands

    A. C. Douma, G. J. Sulter, H. R. Waterham & W. Harder

  2. Laboratory for Electron Microscopy, Biological Centre, University of Groningen, Kerklaan 30, NL-9751 NN, Haren, The Netherlands

    M. Veenhuis

  3. Department of Pharmacology, Catholic University of Leuven, Campus Gasthuisberg, B-3000, Leuven, Belgium

    K. Verheyden & G. P. Mannaerts

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  1. A. C. Douma
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  7. W. Harder
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Douma, A.C., Veenhuis, M., Sulter, G.J. et al. Permeability properties of peroxisomal membranes from yeasts. Arch. Microbiol. 153, 490–495 (1990). https://doi.org/10.1007/BF00248432

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  • DOI: https://doi.org/10.1007/BF00248432

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