Cusabio N-terminal 10xHis-SUMO-tagged Recombinant

Cusabio N-terminal 10xHis-SUMO-tagged Recombinant

February 5, 2022 Off By Bertha Cooper

Properties

Quality level: 200

Recombinant: expressed in E. coli

Rehearsal: ≥95% (SDS PAGE)

Form: lyophilized powder

Molecular weight: 27kDa

Sent in: environment

Storage temperature: −20°C

Related Categories: Proteases

General description

SUMO proteases are a general class of enzymes that specifically remove the protein post-translational modification (PTM) known as a small ubiquitin-related modifier (SUMO), which belongs to the PTM class of ubiquitin and/or ubiquitin-like proteins (UBLs). . The enzyme commonly known as “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 specifically cleaves the SUMO moiety without scarring.

The N-terminal 10xHis-SUMO-tagged Recombinant protease recognizes the ubiquitin-like tertiary structure of the SUMO domain and hydrolyzes the peptide bond in the sequence x–Gly–Gly–x after the Gly-Gly bond, at the C-terminus of the SUMO domain. In addition to cleavage of naturally occurring 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. A histidine tag can also be added at the N-terminus of the SUMO domain as a purification tag.

This SUMO protease product carries a 6x histidine tag. Therefore, it can be easily removed together with the cleaved SUMO domain, after the digestion reaction. SUMO protease is active over a wide range of temperatures (2 to 30°C), ionic strengths (0 to 400 mM NaCl), and pH ranges (6 to 8.5). However, its activity may vary depending on the substrate and conditions. Researchers will need to optimize their specific reaction conditions. As a starting suggestion, 20 SUMO protease units per mg of the target protein can be used for 1 hour at 30°C or overnight at 2–8°C. Cleavage efficiency can then be estimated by SDS-PAGE.

If necessary, the amount of SUMO protease can be adjusted. SUMO protease works best in the presence of reducing agents, eg, 0.5 to 2 mM DTT. DTT in the reaction mix can significantly improve cleavage efficiency, especially during longer incubations. Store reconstituted product at –20°C. It is recommended to reconstitute the enzyme in 100 µL of water or 50% (v/v) glycerol, supplemented with 1 mM DTT. Water/DTT solutions should be stored in frozen aliquots to avoid freeze-thaw cycles, which can negatively affect protease activity.

Unit Definition

One unit of enzyme is defined as the amount that will cut 90% of 100 pmol of SUMO-GST in 1 hour at 30°C