http://www.cnr.it/ontology/cnr/individuo/prodotto/ID45692
Shotgun proteomics for the characterisation of subunit composition of mitochondrial complex I. (Articolo in rivista)
- Type
- Label
- Shotgun proteomics for the characterisation of subunit composition of mitochondrial complex I. (Articolo in rivista) (literal)
- Anno
- 2006-01-01T00:00:00+01:00 (literal)
- Alternative label
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- Pocsfalvi Gabriella; Cuccurullo Manuela; Schlosser Gitta; Cacace Giuseppina; Siciliano Rosa Anna; Mazzeo Maria Fiorella; Scacco Salvatore; Cocco Tiziana; Gnoni Antonio; Malorni Antonio; Papa Sergio (literal)
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- Pagina fine
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- Rivista
- Note
- ISI Web of Science (WOS) (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
- Proteomic and Biomolecular Mass Spectrometry Centre, Institute of Food Science and Technology, CNR, Avellino
Hungarian Academy of Sciences, Eötvös L. University, Budapest
Institute of Medical Biochemistry and Chemistry, University of Bari, Bari
Institute of Biomembranes and Bioenergetics, Italian National Research Council, Bari (literal)
- Titolo
- Shotgun proteomics for the characterisation of subunit composition of mitochondrial complex I. (literal)
- Abstract
- Here we propose shotgun proteomics as an alternative method to gel-based bottom-up proteomic platform for the structural characterization of mitochondrial NADH:ubiquinone oxidoreductase (complex I). The approach is based on simultaneous identification of subunits after global digestion of the intact complex. Resulting mixture of tryptic peptides is purified, concentrated, separated and online analyzed using nano-scale reverse-phase nano-ESI-MS/MS in a single information dependent acquisition mode. The usefulness of the method is demonstrated in our work on the well described model system of complex I from bovine heart mitochondria. The shotgun method led to the identification and partial sequence characterization of 42 subunits representing more than 95% coverage of the complex. In particular, almost all nuclear (except MLRQ) and 5 mitochondria DNA encoded subunits (except ND4L and ND6) were identified. Furthermore, it was possible to identify 30 co-purified proteins of the inner mitochondrial membrane structurally not belonging to complex I. The method's efficiency is shown by comparing it to two classical 1 D gel-based strategies. Shotgun proteomics is less laborious, significantly faster and requires less sample material with minimal treatment, facilitating the screening for post-translational modifications and quantitative and qualitative differences of complex I subunits in genetic disorders. (literal)
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- Autore CNR
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