Table I.

MuRF1 binds a variety of myofibrillar proteins present in muscle homogenate (3,000 g supernatant)

Protein name Gene Gene ID (NCBI) Precipitated from normal muscle with pure MuRF1 Ubiquitylated after addition of E1, E2, and ATP Immunoprecipitated from denervated muscle with Myc-ΔR-MuRF1 
MyBP-C Mybpc2 233199 + + + 
Myosin regulatory light chain (MyLC2) Mylpf 17907 + + + 
Myosin essential light chain (MyLC1) Myl1 17901 + + + 
Myosin heavy chain 1 Myh1 17879 + + + 
Myosin heavy chain 4 Myh4 17884 +  + 
Myosin heavy chain 8 Myh8 17885 +  + 
Myomesin 1 Myom1 17929 +  + 
Actin, alpha skeletal muscle Acta1 11459 + + + 
Filamin C, gamma isoform 1 Flnc 68794 +  + 
Protein name Gene Gene ID (NCBI) Precipitated from normal muscle with pure MuRF1 Ubiquitylated after addition of E1, E2, and ATP Immunoprecipitated from denervated muscle with Myc-ΔR-MuRF1 
MyBP-C Mybpc2 233199 + + + 
Myosin regulatory light chain (MyLC2) Mylpf 17907 + + + 
Myosin essential light chain (MyLC1) Myl1 17901 + + + 
Myosin heavy chain 1 Myh1 17879 + + + 
Myosin heavy chain 4 Myh4 17884 +  + 
Myosin heavy chain 8 Myh8 17885 +  + 
Myomesin 1 Myom1 17929 +  + 
Actin, alpha skeletal muscle Acta1 11459 + + + 
Filamin C, gamma isoform 1 Flnc 68794 +  + 

MuRF1-bound proteins were immunoprecipitated together with MuRF1 from gastrocnemius muscles and identified by mass spectrometry. When a subsequent in vitro ubiquitylation was applied, mass spectrometry analysis identified MyBP-C, MyLC 1 and 2, MyHC, actin, and myomesin as ubiquitylated species.

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