In Vitro Characterization of Chain Depolymerization Activities of SUMO-Specific Proteases.

摘要: SUMO-specific proteases, known as Ulps in baker's yeast and SENPs humans, have important roles controlling the dynamics of SUMO-modified proteins. They display distinct modes action specificity, that they may act on SUMO precursor, mono-sumoylated, and/or polysumoylated proteins, might be specific for substrates with certain paralogs. chains dismantled either by endo or exo mechanisms. Biochemical characterization a protease usually requires purification protein interest. Developing protocol, however, can very difficult, some cases, isolation its pure form go along substantial loss activity. To characterize reaction mechanism Ulps, we developed an vitro assay, which makes use endowed artificial poly-SUMO defined lengths, S. cerevisiae Ulp enzymes crude extract from E. coli. This fast economic approach should applicable to proteases other species well.