Phospho(Ser65)-Ubiquitin Mutant Biotinylated

Ubiquitin is a small polypeptide that can be conjugated via its C-terminus to amine groups of lysine residue on target proteins. Additional ubiquitin moieties can be subsequently conjugated to this initial ubiquitin, utilizing anyone (of seven) lysine residues on the surface of ubiquitin. Biotinylation and phosphorylation were performed on mutant ubiquitin to facilitate biotinylation on amino acid residue 20 that is changed from a serine to a cysteine. The primary function of ubiquitination is to mark proteins for degradation. More specifically, defective mitochondria are selectively degraded after being marked with ubiquitin through a mechanism involving PTEN-induced putative kinase 1 (PINK1) and Parkin. PINK1 is stabilized and accumulates in depolarized mitochondria, where it phosphorylates many proteins. Parkin itself is activated through PINK1-mediated phosphorylated at Ser65 as well as interacting with phosphorylated ubiquitin (also accomplished with PINK1 at Ser65). Phospho-Ubiquitin (pS65) is enzymatically generated with PINK1 kinase from Red Flour Beetle (Tribolium castaneum).

Product Information

Purity	≥ 98% SDS-PAGE and Mass Spectrometry
Molecular Weight	9186 Da
Physical State	Liquid
Quantity	100 µg or 200 µg
Solubility	>30 mg/ml
Formulation	PBS pH 7.4, 5mM TCEP
Storage	-80°C, avoid freeze/thaw cycles