Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF

R&D Systems | Catalog # 11697-SO

His-tag
R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant S. cerevisiae SUMO Protease ULP1 His Protein (11697-SO)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

NP_015305.1

Applications

Enzyme Activity
View all SUMO Protease ULP1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived yeast SUMO Protease ULP1 protein
Leu403-Lys621, with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

26 kDa

SDS-PAGE

27-33 kDa, under reducing conditions.

Activity

Measured by its ability to cleave SUMO-GFP Protease Substrate.
The DC50 is <0.8 ng, as measured under the described conditions.The DC50 is defined as the amount of enzyme required to cleave 50% of 1 μg SUMO-GFP in 2 hours at room temperature. Use of Recombinant Yeast SUMO Protease ULP1 in the cleavage of other protein substrates may require alternative conditions for optimal performance.

Scientific Data Images for Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF

Recombinant Yeast SUMO Protease ULP1 His-tag Protein Enzyme Activity.

Recombinant Yeast SUMO Protease ULP1 His-tag activity is measured by its ability to cleave SUMO-GFP Protease Substrate (11698-SO). The R&D Systems protein has higher activity than competitor protein in direct comparison.

Recombinant Yeast SUMO Protease ULP1 His-tag Protein Enzyme Activity.

Recombinant Yeast SUMO Protease ULP1 His-tag Protein (Catalog # 11697-SO) from R&D Systems and a competitor product are able to cleave Recombinant SUMO-GFP Protease Substrate (~47 kDa, 11698-SO) per insert assay conditions. Cleavage is detected with SDS-PAGE under reducing (R) conditions and visualized by Coomassie® Blue staining. The enzyme from R&D Systems offers a better value than the competition.

Recombinant Yeast SUMO Protease ULP1 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Yeast SUMO Protease ULP1 His-tag Protein (Catalog # 11697-SO) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 27-33 kDa, under reducing conditions.

Formulation, Preparation, and Storage

11697-SO
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: SUMO Protease ULP1

Recombinant Yeast SUMO Protease ULP1 (Ubiquitin-like-specific protease 1), commonly known as SUMO protease 1, is essential in the processing and deconjugation of a small ubiquitin-like modifier (SUMO), SMT3, from targeted proteins in the SUMO pathway of yeast where it plays an essential role in the cell cycle (1, 2). ULP1 is a 621 amino acid protein from the peptidase C48 family of cysteine proteases and contains a conserved C-terminal protease fold domain and a weakly conserved N-terminal domain (1, 3). The catalytic site with a conserved cysteine protease structure and catalytic triad is located in a shallow and narrow cleft that creates limited access to the active site through the constricted hydrophobic tunnel and confers high specificity to the enzyme (3). ULP1 cleaves the C-terminus of SUMO to a mature form after a glycine-glycine pair. Use of SUMO as a fusion for recombinant proteins with subsequent processing by ULP1 in E. coli was explored due to the demand to find improved methods for recombinant production of proteins for use in academia and industry (4). SUMO as a fusion partner was found to enhance expression in E. coli for a diverse population of protein targets and unlike other common fusion partners SUMO could serve as a chaperone for correct folding such that the target would remain soluble after the SUMO protein was removed (4). In addition, ULP1 was found to be capable of cleaving with high efficiency a range of targets specifically at the SUMO site under a broad variety of reaction conditions (5, 6). Use of SUMO fusion with ULP1 cleavage is a tool that allows for successful recombinant protein production with a native N-terminal sequence (5) providing significant benefits over other fusion and cleavage methods being used (4,7-10). Use of recombinant SUMO fusions with subsequent ULP1 cleavage is a beneficial tool for recombinant protein production in both academic research and industrial biotechnology (4, 5, 11).

References

  1. Li, S.J and M. Hochstrasser (1999) Nature 398:246.
  2. Li, S.J and M. Hochstrasser (2000) Mol Cell Biol. 20:2367.
  3. Mossessova, E. and C.D. Lima (2000) Mol. Cell 5:865.  
  4. Malakhov, M.P. et al. (2004) J. Struct. Funct. Genomics 5:75.  
  5. Marblestone, J.G. et al. (2006) Protein Sci. 15:182.
  6. Satakarni, M. and R. Curtis (2011) Protein Expr. Purif. 78:113.
  7. Baker, R.T. (1996) Curr. Opin. Biotechnol. 7:541.
  8. Jonasson, P. et al. (2002) Biotechnol. Appl. Biochem. 35:91.
  9. Butt, T.R. et al. (2005) Protein Expr. Purif. 43:1.
  10. Peroutka, R.J. et al. (2011) Methods Mol. Biol. 705:15.
  11. Wang, Z. et al. (2010) Protein Expr. Purif. 73:203.

Alternate Names

EC 3.4.22.68, NIB1

Entrez Gene IDs

856087 (Yeast)

Gene Symbol

ULP1

UniProt

NP_015305.1

Additional SUMO Protease ULP1 Products

Product Documents for Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF

For research use only

Customer Reviews for Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF and earn rewards!

Have you used Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant S. cerevisiae SUMO Protease ULP1 His Protein, CF (11697-SO):

Materials
  • Assay Buffer: 50 mM Tris, 5 mM DTT, pH 8.0 
  • Recombinant Yeast SUMO Protease ULP1 His-tag (SUMO Protease ULP1) (Catalog # 11697-SO)
  • Recombinant SUMO-GFP Protease Substrate (rSUMO-GFP) (Catalog # 11698-SO)
  • 15% SDS-PAGE gel
  • Reducing Sample Buffer
  • Gel Staining Reagent
  1. Prepare a curve of SUMO Protease ULP1 by diluting SUMO Protease ULP1 to 20, 1.25, 0.312, 0.156, 0.078, 0.039 and 0.00977 µg/mL in Assay Buffer.
  2. Dilute rSUMO-GFP to 200 µg/mL in Assay Buffer.
  3. Combine 20 µL of each SUMO Protease ULP1 curve dilution and 20 µL of 200 µg/mL rSUMO-GFP.  Prepare a Control by combining 20 µL of 200 µg/mL rSUMO-GFP and 20 µL Assay Buffer.
  4. Incubate reaction mixtures and control at room temperature for 2 hours.
  5. Stop the reactions by combining equal volumes of reaction mixture (including control) and Reducing SDS-PAGE Sample Buffer. Heat all samples at 95 °C for 3 minutes.
  6. Load 20 µL of each stopped reaction per lane on a 15% SDS-PAGE gel and perform electrophoresis.
  7. Stain gel and analyze the % cleavage of rSUMO-GFP using densitometry for each SUMO Protease ULP1 curve dilution.  
  8. Determine the DC50 by plotting % cleavage vs SUMO Protease ULP1 concentration (ng) using 4-PL fitting.
Per Lane:
  • SUMO Protease ULP1: 100, 6.25, 1.56, 0.78, 0.39, 0.195, 0.0488, and 0 ng
  • rSUMO-GFP: 1 µg

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes