title: Focused proteomics: towards a high throughput monoclonal antibody-based resolution of proteins for diagnosis of mitochondrial diseases.
abstract: The availability of monoclonal antibodies (mAbs) against the proteins of the oxidative phosphorylation chain (OXPHOS) and other mitochondrial components facilitates the analysis and ultimately the diagnosis of mitochondrially related diseases. mAbs against each of the five complexes and pyruvate dehydrogenase (PDH) are the basis of a rapid and simple immunocytochemical approach [Hanson, B.J., Capaldi, R.A., Marusich, M.F. and Sherwood, S.W., J. Histochem. Cytochem. 50 (2002) 1281-1288]. This approach can be used to detect if complexes have altered assembly in mitochondrial disease due to mutations in nuclear encoded genes, such as in Leigh's disease, or in mitochondrially encoded genes, e.g., MELAS. Other mAbs have recently been obtained that can immunocapture each of the five OXPHOS complexes, PDH and the adenine nucleotide translocase (ANT) from very small amounts of tissue such as that obtained from cell culture or needle biopsies from patients. When adapted to a 96-well plate format, these mAbs allow measurement of the specific activity of each of the mitochondrial components individually and analysis of their subunit composition and state of posttranslational modification. The immunocapture protocol should be useful not only in the analysis of genetic mitochondrial diseases but also in evaluating and ultimately diagnosing late-onset mitochondrial disorders including Parkinson's disease, Alzheimer's disease, and late-onset diabetes, which are thought to result from accumulated oxidative damage to mitochondrial proteins such as the OXPHOS chain.
authors.affiliation: Institute of Molecular Biology, Department of Biology, University of Oregon, Eugene OR 97403-1229, USA.
authors.forename: James
authors.initials: J
authors.lastname: Murray
authors.forename: Sky
authors.initials: S
authors.lastname: Yonally
authors.forename: Robert
authors.initials: R
authors.lastname: Aggeler
authors.forename: Michael F
authors.initials: MF
authors.lastname: Marusich
authors.forename: Roderick A
authors.initials: RA
authors.lastname: Capaldi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D003576:Electron Transport Complex IV / Q000032:analysis / Q000378:metabolism; D005347:Fibroblasts / Q000276:immunology / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D000226:Mitochondrial ADP, ATP Translocases / Q000032:analysis / Q000276:immunology; D028361:Mitochondrial Diseases / Q000175:diagnosis* / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D011768:Pyruvate Dehydrogenase Complex / Q000032:analysis / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011506:Proteins; D011768:Pyruvate Dehydrogenase Complex; D000226:Mitochondrial ADP, ATP Translocases; D003576:Electron Transport Complex IV