Titre : | An integrated protein biomarker discovery and identification pipeline for investigating the skeletal muscle proteome (abstract : congrès international de Myologie, 2005) |
contenu dans : | |
Auteurs : | Congrès international de myologie 2005 (International Congress of Myology 2005; 9-13 mai 2005; Nantes, France) ; Le Bihan M ; Hou Y ; Coulton G |
Type de document : | Article |
Année de publication : | 2005 |
Pages : | p. 91 |
Langues: | Anglais |
Mots-clés : | colloque ; culture cellulaire ; cyphoscoliose ; dégenérescence de la fibre musculaire ; muscle squelettique ; protéome ; régénération musculaire ; souris |
Résumé : |
Communication n° 356. Introduction : Skeletal muscle can hypertrophy, atrophy and alter fibre type in response to external stimuli. Disease also generates typical pathologic changes in phenotype. Identification of molecular networks underpinning such phenotypic change will be the basis for new treatments for muscle disease. Objective : Over the last year we have developed a robust, and sensitive analytical pipeline for characterising global changes in muscle proteomes both from whole muscle and cell culture. Methods : The method is based upon the integrated application of Ciphergen protein arrays, Surface-Enhanced Laser Desorption and Ionisation (SELDI) mass spectrometry, 2D gel electrophoresis and Matrix Assisted Laser Desorption and Ionisation (MALDI) mass spectrometry. The strategy comprises several stages. 1) Protein extraction, 2) Biomarker Discovery, 3) Biomarker validation & 4) Biomarker Identification. Results : We have employed this strategy in a proof-of-principle model system namely the identification of phenotypic differences between fast and slow mouse muscles. Secondarily we have analysed protein changes in muscles from kyphoscoliosis (ky) mutant mice. Ky muscles exhibit a profound neonatal degenerative myopathy. Following regeneration their muscles cannot hypertrophy but do show profound shifts to slow muscle fibre type (adult ky soleus = 100% type 1 fibres). Using this technique, more than 200 proteins were detected below 50kD in skeletal muscle. Sixty of which were statistically validated protein biomarkers discerning fast from slow twitch and normal from ky skeletal muscle. Eight of them were formally identified. Conclusions : We show that this integrated proteomic pipeline is reliable, robust and rapid and has the potential to make a significant contribution to the discovery and identification of protein biomarkers characteristic of normal adaptation and also diagnostic of pathological change in skeletal muscle. |