BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1
Protein degradation is essential for maintaining cellular homeostasis. The proteasome is the centralenzyme responsible for non-lysosomal protein degradation in eukaryotic cells. Although proteasome assembly is not yet completelyunderstood, a number of cofactors required for proper assembly and matur...
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| Format: | Article |
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Elsevier
2013-04-01
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| Series: | Computational and Structural Biotechnology Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037014600295 |
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| author | Bebiana Sá-Moura Ana Marisa Simões Joana Fraga Humberto Fernandes Isabel A. Abreu Hugo M. Botelho Cláudio M. Gomes António J. Marques R. Jürgen Dohmen Paula C. Ramos Sandra Macedo-Ribeiro |
| author_facet | Bebiana Sá-Moura Ana Marisa Simões Joana Fraga Humberto Fernandes Isabel A. Abreu Hugo M. Botelho Cláudio M. Gomes António J. Marques R. Jürgen Dohmen Paula C. Ramos Sandra Macedo-Ribeiro |
| author_sort | Bebiana Sá-Moura |
| collection | DOAJ |
| description | Protein degradation is essential for maintaining cellular homeostasis. The proteasome is the centralenzyme responsible for non-lysosomal protein degradation in eukaryotic cells. Although proteasome assembly is not yet completelyunderstood, a number of cofactors required for proper assembly and maturation have been identified. Ump1 is a short-livedmaturation factor required for the efficient biogenesis of the 20S proteasome. Upon the association of the two precursorcomplexes, Ump1 is encased and is rapidly degraded after the proteolytic sites in the interior of the nascent proteasome areactivated. In order to further understand the mechanisms behind proteasomal maturation, we expressed and purified yeast Ump1 inE. coli for biophysical and structural analysis.
We show that recombinant Ump1 is purified as a mixture of different oligomeric species and thatoligomerization is mediated by intermolecular disulfide bond formation involving the only cysteine residue present in the protein.Furthermore, a combination of bioinformatic, biochemical and structural analysis revealed that Ump1 shows characteristics of anintrinsically disordered protein, which might become structured only upon interaction with the proteasomesubunits. |
| first_indexed | 2024-04-11T22:57:00Z |
| format | Article |
| id | doaj.art-0ef0dca3b6c44117b9015a89ce3415b8 |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-04-11T22:57:00Z |
| publishDate | 2013-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-0ef0dca3b6c44117b9015a89ce3415b82022-12-22T03:58:20ZengElsevierComputational and Structural Biotechnology Journal2001-03702013-04-017810.5936/csbj.201304006BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1Bebiana Sá-Moura0Ana Marisa Simões1Joana Fraga2Humberto Fernandes3Isabel A. Abreu4Hugo M. Botelho5Cláudio M. Gomes6António J. Marques7R. Jürgen Dohmen8Paula C. Ramos9Sandra Macedo-Ribeiro10IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto,PortugalIBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto,PortugalIBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto,PortugalCentre for Molecular and Structural Biomedicine, CBME/IBB, LA, PortugalIBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto,PortugalInstituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, PortugalInstituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, PortugalCentre for Molecular and Structural Biomedicine, CBME/IBB, LA, PortugalInstitute for Genetics, University of Cologne, Zülpicher Str. 47, D-50674 Cologne, GermanyCentre for Molecular and Structural Biomedicine, CBME/IBB, LA, PortugalIBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto,PortugalProtein degradation is essential for maintaining cellular homeostasis. The proteasome is the centralenzyme responsible for non-lysosomal protein degradation in eukaryotic cells. Although proteasome assembly is not yet completelyunderstood, a number of cofactors required for proper assembly and maturation have been identified. Ump1 is a short-livedmaturation factor required for the efficient biogenesis of the 20S proteasome. Upon the association of the two precursorcomplexes, Ump1 is encased and is rapidly degraded after the proteolytic sites in the interior of the nascent proteasome areactivated. In order to further understand the mechanisms behind proteasomal maturation, we expressed and purified yeast Ump1 inE. coli for biophysical and structural analysis. We show that recombinant Ump1 is purified as a mixture of different oligomeric species and thatoligomerization is mediated by intermolecular disulfide bond formation involving the only cysteine residue present in the protein.Furthermore, a combination of bioinformatic, biochemical and structural analysis revealed that Ump1 shows characteristics of anintrinsically disordered protein, which might become structured only upon interaction with the proteasomesubunits.http://www.sciencedirect.com/science/article/pii/S2001037014600295Circular dichroismprotein structuredynamic light scatteringintrinsically disordered |
| spellingShingle | Bebiana Sá-Moura Ana Marisa Simões Joana Fraga Humberto Fernandes Isabel A. Abreu Hugo M. Botelho Cláudio M. Gomes António J. Marques R. Jürgen Dohmen Paula C. Ramos Sandra Macedo-Ribeiro BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 Computational and Structural Biotechnology Journal Circular dichroism protein structure dynamic light scattering intrinsically disordered |
| title | BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 |
| title_full | BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 |
| title_fullStr | BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 |
| title_full_unstemmed | BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 |
| title_short | BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF RECOMBINANT YEAST PROTEASOME MATURATION FACTOR UMP1 |
| title_sort | biochemical and biophysical characterization of recombinant yeast proteasome maturation factor ump1 |
| topic | Circular dichroism protein structure dynamic light scattering intrinsically disordered |
| url | http://www.sciencedirect.com/science/article/pii/S2001037014600295 |
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