Document Detail

Patient-specific protein aggregates in myofibrillar myopathies: Laser microdissection and differential proteomics for identification of plaque components.
MedLine Citation:
PMID:  23044792     Owner:  NLM     Status:  Publisher    
Myofibrillar myopathies (MFM) are histopathologically characterized by desmin-positive protein aggregates and myofibrillar degeneration. While about half of all MFM are caused by mutations in genes encoding sarcomeric and extra-sarcomeric proteins (desmin, filamin C, plectin, VCP, FHL1, ZASP, myotilin, αB-crystallin, and BAG3), the other half of these diseases is due to still unresolved gene defects. The present study aims at the proteomic characterization of pathological protein aggregates in skeletal muscle biopsies from patients with MFM-causing gene mutations. The technical strategy is based on the dissection of plaque- vs. plaque-free tissue areas from the same individual patient by laser dissection microscopy, filter-aided sample preparation, iTRAQ-labeling and analysis on the peptide level using offline nano-LC and MALDI-TOF-TOF tandem mass spectrometry for protein identification and quantification. The outlined workflow overcomes limitations of merely qualitative analyses, which cannot discriminate contaminating non-aggregated proteins. Dependent on the MFM causing mutation different sets of proteins were revealed as genuine (accumulated) plaque components in independent technical replicates: (1) αB-crystallin, desmin, filamin A/C, myotilin, PRAF3, RTN1, SQSTM, XIRP1 and XIRP2 (patient with defined MFM mutation distinct from FHL1) or (2) desmin, FHL1, filamin A/C, KBTBD10, NRAP, SQSTM, RL40, XIRP1 and XIRP2 (patient with FHL1 mutation). The results from differential proteomics indicate that plaques from different patients exhibit protein compositions with partial overlap, on the one hand, and mutation-dependent protein contents on the other. The FHL1 mutation-specific pattern was validated for four patients with respect to desmin, SQSTM, and FHL1 by immunohisto- chemistry.
Sarah Feldkirchner; Joachim Schessl; Stefan Müller; Benedikt Schoser; Franz-Georg Hanisch
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-9
Journal Detail:
Title:  Proteomics     Volume:  -     ISSN:  1615-9861     ISO Abbreviation:  Proteomics     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-9     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Friedrich-Baur-Institute, Dept. of Neurology, Ludwig-Maximilians-University, Munich.
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