Table 1.

Trim32 binds thin filament and Z-band components in the muscle homogenate

Protein name Gene No. of unique peptides (found initially) Precipitated from normal muscle with pure Trim32 Ubiquitylated after the addition of E1, E2, and ATP 
α-Actin Acta1 
α-Actinin Actn3 
Tropomyosin 1 alpha chain Tpm1 
Desmin Des 
Myosin regulatory light chain Mylpf  
Protein name Gene No. of unique peptides (found initially) Precipitated from normal muscle with pure Trim32 Ubiquitylated after the addition of E1, E2, and ATP 
α-Actin Acta1 
α-Actinin Actn3 
Tropomyosin 1 alpha chain Tpm1 
Desmin Des 
Myosin regulatory light chain Mylpf  

Trim32-bound proteins were precipitated together with Trim32 from the soluble fraction of TA and identified by mass spectrometry. When a subsequent in vitro ubiquitylation reaction was carried out, mass spectrometry analysis identified actin, α-actinin, Tm, and desmin as ubiquitylated species.

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