• Purity

  >90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie? Blue stain.

• Activity

  Add Recombinant Human HA-SUMO3 Vinyl Sulfone to     in vitro assays to inhibit SUMO-specific isopeptidases (SENPs). The HA-tag allows for detection and purification of SUMO-specific isopeptidases (SENPs) activity. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HA-SUMO3 Vinyl Sulfone concentration of 1-5 μM.

• Source

  E. coli-derived Contains an N?terminal HA (YPYDVPDYA) tag and a C?terminal Vinyl Sulfone

• Accession #

• Predicted Molecular Mass

  12 kDa

Background: SUMO3

Human Small Ubiquitin-like Modifier 3 (SUMO3), also known as SMT3A, is synthesized as a 103 amino acid (aa), propeptide with a predicted 11.5 kDa. SUMO3 contains a two aa C-terminal prosegment. Human SUMO3 shares 83% sequence identity with mouse SUMO3. SUMO3 also has high aa sequence homology to SUMO2 and SUMO4, 87% and 75%, respectively. SUMO3 shares only 47% sequence identity with SUMO1. SUMOs are a family of small, related proteins that can be enzymatically attached to a target protein by a post-translational modification process termed SUMOylation (1-3). All SUMO proteins share a conserved Ubiquitin domain and a C-terminal diglycine cleavage/attachment site. Following prosegment cleavage, the C-terminal glycine residue of SUMO3 is enzymatically attached to a lysine residue on a target protein. In humans, SUMO3 is conjugated to a variety of molecules in the presence of the SAE1/UBA2 SUMO-activating (E1) enzyme and the UBE2I/Ubc9 SUMO-conjugating (E2) enzyme (4,5). In yeast, the SUMO-activating (E1) enzyme is Aos1/Uba2p (6). Because of the high level of sequence homology most studies report effects of SUMO2/3. For example, addition of SUMO2/3 was shown to modulate the function of ARHGAP21, a RhoGAP protein known to be involved in cell migration (7). Other reports indicate that the conjugation by SUMO2/3, but not SUMO1, may represent an important mechanism to protect neurons during episodes of cerebral ischemia (8,9). However, studies suggest that SUMO2/3 expression is regulated in an isoform-specific manner since oxidative stress downregulated the transcription of SUMO3 but not SUMO2 (10).

This N-terminal HA-tagged SUMO is a potent, irreversible, and specific inhibitor of SUMO-specific proteases (SENPs). This protein inhibits the hydrolysis of poly-SUMO chains on substrate proteins in vitro and thus enhances poly-SUMO chain accumulation. The HA peptide sequence (YPYDVPDYA) is derived from the influenza Hemagglutinin protein. This epitope allows for the sensitive identification or purification of SENP activities since it is specifically recognized by anti-HA antibodies and/or anti-HA-agarose.

• References:

1. Desterro, J.M. et al. (1997) FEBs. Lett. 417:297.

2. Bettermann, K. et al. (2012) Cancer Lett. 316:113.

3. Praefcke, G.J. et al. (2012) Trends Biochem. Sci. 37:23.

4. Okuma, T. et al. (1999) Biochem. Biophys. Res. Commun. 254:693.

5. Tatham, M.H. et al. (2001) J. Biol. Chem. 276:35368.

6. Johnson, E.S. et al. (1997) EMBO J. 16:5509.

7. Bigarella, C.L. et al. (2012) FEBS Lett. 586:3522.

8. Datwyler, A.L. et al. (2012) J. Cereb. Blood Flow Metab. 31:2152.

9. Wang, Z. et al. (2012) Protein Expr. Purif. 82:174.

10. Sang, J. et al. (2012) Biochem. J. 435:489.

• Long Name:

  Small Ubiquitin-like Modifier 3

• Entrez Gene IDs:

  6612 (Human)

• Alternate Names:

  SMT3 (suppressor of mif two 3, yeast) homolog 1; SMT3 homolog 1; SMT3 suppressor of mif two 3 homolog 1; SMT3 suppressor of mif two 3 homolog 3 (S. cerevisiae); SMT3 suppressor of mif two 3 homolog 3 (yeast); SMT3A; SMT3B; SMT3H1; SMT3H1small ubiquitin-related modifier 3; SUMO-2; SUMO3; SUMO-3; ubiquitin-like protein SMT3A; Ubiquitin-like protein SMT3B