Phosphoproteomic analysis of aged skeletal muscle.
One of the most important post-translational modifications is represented by phosphorylation on tyrosine, threonine and serine residues. Since abnormal phosphorylation is associated with various pathologies, it was of interest to perform a phosphoproteomic profiling of age-related skeletal muscle degeneration. We used the fluorescent phospho-specific Pro-Q Diamond dye to determine whether changes in the overall phosphorylation of the soluble skeletal muscle proteome differs significantly between young adult and senescent fibres. As an established model system of sarcopenia, we employed 30-month-old rat gastrocnemius fibres. Following the mass spectrometric identification of 59 major 2-D phosphoprotein landmark spots, the fluorescent dye staining survey revealed that 22 muscle proteins showed a differential expression pattern between 3-month- and 30-month-old muscle. Increased phosphorylation levels were shown for myosin light chain 2, tropomyosin alpha, lactate dehydrogenase, desmin, actin, albumin and aconitase. In contrast, decreased phospho-specific dye binding was observed for cytochrome c oxidase, creatine kinase and enolase. Thus, aging-induced alterations in phosphoproteins appear to involve the contractile machinery and the cytoskeleton, as well as the cytosolic and mitochondrial metabolism. This confirms that sarcopenia of old age is a complex neuromuscular pathology that is associated with drastic changes in the abundance and structure of key muscle proteins.
Gannon
Joan
J
Department of Biology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Staunton
Lisa
L
O'Connell
Kathleen
K
Doran
Philip
P
Ohlendieck
Kay
K
D000375:Aging / Q000378:metabolism*; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005453:Fluorescence; D009124:Muscle Proteins / Q000032:analysis; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis*; D010766:Phosphorylation; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D051381:Rats; D017208:Rats, Wistar; D013194:Staining and Labeling
D009124:Muscle Proteins; D010750:Phosphoproteins; D020543:Proteome