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J. Biol. Chem., Vol. 282, Issue 43, 31267-31272, October 26, 2007

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Ubiquitination of Mammalian Pex5p, the Peroxisomal Import Receptor^*

Andreia F. Carvalho¹, Manuel P. Pinto¹, Cláudia P. Grou¹, Inês S. Alencastre, Marc Fransen^¶, Clara Sá-Miranda, and Jorge E. Azevedo²

From the Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal, the Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Largo Professor Abel Salazar, 2, 4009-003 Porto, Portugal, and the ^¶Faculteit Geneeskunde, Departement Moleculaire Celbiologie, Katholieke Universiteit Leuven, Herestraat 49, B-3000 Leuven, Belgium

Protein translocation across the peroxisomal membrane requires the concerted action of numerous peroxins. One central component of this machinery is Pex5p, the cycling receptor for matrix proteins. Pex5p recognizes newly synthesized proteins in the cytosol and promotes their translocation across the peroxisomal membrane. After this translocation step, Pex5p is recycled back into the cytosol to start a new protein transport cycle. Here, we show that mammalian Pex5p is ubiquitinated at the peroxisomal membrane. Two different types of ubiquitination were detected, one of which is thiol-sensitive, involves Cys¹¹ of Pex5p, and is necessary for the export of the receptor back into the cytosol. Together with mechanistic data recently described for yeast Pex5p, these findings provide strong evidence for the existence of Pex4p- and Pex22p-like proteins in mammals.

Received for publication, June 23, 2007 , and in revised form, August 27, 2007.

^* This work was supported in part by the POCTI Program of the Fundação para a Ciência e Tecnologia; the Fundo Europeu de Desenvolvimento Regional, Portugal; and European Union VI Framework Program Grant LSHG-CT-2004-512018 (Peroxisomes in Health and Disease). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by the Fundação para a Ciência e Tecnologia.

² To whom correspondence should be addressed: Inst. de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal. Tel.: 351-226-074-900; Fax: 351–226-099-157; E-mail: jazevedo{at}ibmc.up.pt.

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