Hyperubiquitination of proteins in dilated cardiomyopathy.
Protein degradation by the ubiquitin-proteasome pathway plays an important role in a variety of fundamental cellular processes, including cell cycle regulation, transcription, antigen processing and muscle remodelling. Research into disorders associated with the ubiquitin-proteasome system has been...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2003
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| _version_ | 1797094229451735040 |
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| author | Weekes, J Morrison, K Mullen, A Wait, R Barton, P Dunn, M |
| author_facet | Weekes, J Morrison, K Mullen, A Wait, R Barton, P Dunn, M |
| author_sort | Weekes, J |
| collection | OXFORD |
| description | Protein degradation by the ubiquitin-proteasome pathway plays an important role in a variety of fundamental cellular processes, including cell cycle regulation, transcription, antigen processing and muscle remodelling. Research into disorders associated with the ubiquitin-proteasome system has been mainly in the field of neurodegenerative diseases. It is however becoming increasingly apparent that defects in the system are responsible for a number of non-neurological pathologies. Based on initial observations made as part of a proteomic analysis of an animal model of dilated cardiomyopathy (DCM) which indicated increased activity of the ubiquitin-proteasome system, we sought to determine whether this system was perturbed in hearts of human DCM patients. We studied explanted hearts from 12 DCM, 9 ischaemic (IHD) and 12 unused donor hearts. Protein expression was examined using two-dimensional polyacrylamide gel electrophoresis, Western blotting and immunohistochemistry. Expression of mRNA was examined using real-time quantitative polymerase chain reaction. Ubiquitinated proteins were affinity purified using a ubiquitin-binding column and identified using peptide mass fingerprinting. All DCM hearts showed significantly higher expression of certain key enzymes of the ubiquitin-proteasome pathway. mRNA expression of ubiquitin carboxyl-terminal hydrolase (UCH) was significantly higher (5.4-fold) in DCM hearts than in control hearts. Myocytes in sections from DCM hearts stained positively for UCH, whereas control hearts were negative. Overall protein ubiquitination was increased two-fold in DCM relative to IHD hearts and five-fold relative to donor hearts. The ubiquitination of a number of distinct proteins was greatly enhanced in DCM hearts as revealed by anti-ubiquitin Western blots. A number of these proteins were identified using affinity purification and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. |
| first_indexed | 2024-03-07T04:11:12Z |
| format | Journal article |
| id | oxford-uuid:c7dee4a8-7ada-4763-aed9-a54ac612d224 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T04:11:12Z |
| publishDate | 2003 |
| record_format | dspace |
| spelling | oxford-uuid:c7dee4a8-7ada-4763-aed9-a54ac612d2242022-03-27T06:48:26ZHyperubiquitination of proteins in dilated cardiomyopathy.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c7dee4a8-7ada-4763-aed9-a54ac612d224EnglishSymplectic Elements at Oxford2003Weekes, JMorrison, KMullen, AWait, RBarton, PDunn, MProtein degradation by the ubiquitin-proteasome pathway plays an important role in a variety of fundamental cellular processes, including cell cycle regulation, transcription, antigen processing and muscle remodelling. Research into disorders associated with the ubiquitin-proteasome system has been mainly in the field of neurodegenerative diseases. It is however becoming increasingly apparent that defects in the system are responsible for a number of non-neurological pathologies. Based on initial observations made as part of a proteomic analysis of an animal model of dilated cardiomyopathy (DCM) which indicated increased activity of the ubiquitin-proteasome system, we sought to determine whether this system was perturbed in hearts of human DCM patients. We studied explanted hearts from 12 DCM, 9 ischaemic (IHD) and 12 unused donor hearts. Protein expression was examined using two-dimensional polyacrylamide gel electrophoresis, Western blotting and immunohistochemistry. Expression of mRNA was examined using real-time quantitative polymerase chain reaction. Ubiquitinated proteins were affinity purified using a ubiquitin-binding column and identified using peptide mass fingerprinting. All DCM hearts showed significantly higher expression of certain key enzymes of the ubiquitin-proteasome pathway. mRNA expression of ubiquitin carboxyl-terminal hydrolase (UCH) was significantly higher (5.4-fold) in DCM hearts than in control hearts. Myocytes in sections from DCM hearts stained positively for UCH, whereas control hearts were negative. Overall protein ubiquitination was increased two-fold in DCM relative to IHD hearts and five-fold relative to donor hearts. The ubiquitination of a number of distinct proteins was greatly enhanced in DCM hearts as revealed by anti-ubiquitin Western blots. A number of these proteins were identified using affinity purification and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. |
| spellingShingle | Weekes, J Morrison, K Mullen, A Wait, R Barton, P Dunn, M Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title | Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title_full | Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title_fullStr | Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title_full_unstemmed | Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title_short | Hyperubiquitination of proteins in dilated cardiomyopathy. |
| title_sort | hyperubiquitination of proteins in dilated cardiomyopathy |
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