General description
SUMO proteases are enzymes that specifically cleave the post-translational protein modification (PTM) known as small ubiquitin-related modifier (SUMO). SUMO falls into the PTM class of ubiquitin and/or ubiquitin-like proteins (UBL).SUMO protease is the Ubl-specific protease 1 (Ulp1) from Saccharomyces cerevisiae. This was the first of this class of enzymes to be isolated. SUMO protease cleaves specifically the SUMO moiety in a ‘scarless′ manner. After recognizing the tertiary structure of the Ubiquitin-like SUMO domain, SUMO protease hydrolyzes the peptide bond in the x–Gly–Gly–x sequence after the Gly-Gly bond, at the C-terminus of the SUMO domain. Besides the cleavage of natural SUMO-modified proteins, SUMO protease is used to cleave recombinant SUMO fusion proteins. The SUMO domain is a known solubility-enhancing fusion tag used in recombinant protein expression. Since this recombinant protease does not contain any coman protein purification tag it can be used for on-column cleavage of column bound SUMO fusion protein. Sumo protease with Biotin tag can be easily removed at the end of the digestion reaction.
Application
This biotin-tagged SUMO protease product is designed to be used for on-column cleavage of SUMO fusion proteins. This method specifically cleaves the protein of interest from a column-bound SUMO fusion protein, leaving the SUMO domain bound to the affinity column (e.g. Ni-NTA column) and eluting only the protein of interest. This method is advantageous to post-elution cleavage for several reasons:
Eliminates most of the impurities normally associated with purification on Ni-chelating columns.
- Allows much gentler elution conditions, with an added flexibility in the composition of the elution buffer.
- Assist preventing protein aggregation and inactivation.
- Following cleavage, the protease can be efficiently removed by using any avidin-conjugated or streptavidin-conjugated beads.
This biotin-tagged SUMO protease has been enzymatically biotinylated without affecting its proteolytic activity. It does not include any additional protein purification tag (e.g., histidine-tag or GST).
Other Notes
One enzyme unit is defined as the amount that will cut 90% of 100 pmol of SUMO-GST in 1 hour at 30°C.
| recombinant | expressed in E. coli |
| Quality Segment | 200 |
| assay | ≥90% (SDS-PAGE) |
| form | aqueous solution |
| mol wt | 27 kDa |
| concentration | ≥25,000 units/mL |
| shipped in | dry ice |
| storage temp. | −20°C |
