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Originally published In Press as doi:10.1074/jbc.M004366200 on July 10, 2000

J. Biol. Chem., Vol. 275, Issue 42, 32444-32451, October 20, 2000
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Characterization of Peroxisomal Pex5p from Rat Liver
Pex5p IN THE Pex5p-Pex14p MEMBRANE COMPLEX IS A TRANSMEMBRANE PROTEIN*

Alexandra M. M. GouveiaDagger §, Carlos ReguengaDagger §, Márcia E. M. OliveiraDagger §, Clara Sá-MirandaDagger ||, and Jorge E. AzevedoDagger §**

From the Dagger  Instituto de Biologia Molecular e Celular and § Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 4150-180 Porto and || Instituto de Genética Médica Jacinto de Magalhães, 4050-466 Porto, Portugal

Pex5p is the receptor for the vast majority of peroxisomal matrix proteins. Here, we show that about 15% of rat liver Pex5p is found in the peroxisomal fraction representing 0.06% of total peroxisomal protein. This population of Pex5p displays all the characteristics of an intrinsic membrane protein. Protease protection assays indicate that this pool of Pex5p has domains exposed on both sides of the peroxisomal membrane. The strong interaction of Pex5p with the membrane of the organelle is not affected by mild protease treatment of intact organelles, conditions that result in the partial degradation of Pex13p. Cytosolic Pex5p is a monomeric protein. In contrast, virtually all peroxisomal Pex5p was found to be part of a stable 250-kDa protein assembly. This complex was isolated and shown to comprise just two subunits, Pex5p and Pex14p.

* This work was supported in part by Ministério da Ciência e Tecnologia, Portugal, Grant PRAXIS XXI 2/2.1/SAU/1345/95.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by Ministério da Ciência e Tecnologia, Portugal.

** To whom correspondence should be addressed: IBMC-UP, Rua do Campo Alegre 823, 4150-180 Porto, Portugal. Tel.: 351-226074900; Fax: 351-226092404; E-mail: jazevedo@ibmc.up.pt.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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