SUMO1 [6His-tagged]

Ubiquigent

Catalog No: UBI-60-0002-500

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Species Reactivity: Human 
Source: E. coli expression 
Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol  
Concentration: 1 mg/ml 
Molecular Weight: ~13.5 kDa  
Protein Sequence: Accession number: NP_003343. For full protein sequence information download the Certificate of Analysis pdf.  
Quality Control Protein Identification: Confirmed by mass spectrometry.  
Quality Control Activity: E2 Thioester SUMO-1 Loading Assay:  The activity of His-SUMO-1 was validated by loading His-SUMO-1 onto the active cysteine of the UBE2I E2 enzyme via a transthiolation reaction. Incubation of His-SUMO-1, SAE1/SAE2 and UBE2I enzymes in the presence of ATP at 30 °C was compared at two time points, T0 and T10 minutes. Sensitivity of the His-SUMO-1/UBE2I thioester bond to the reducing agent DTT was confirmed.  

Background

Small Ubiquitin-like Modifiers (SUMOs) are a family of small, related proteins that can be enzymatically conjugated to a target protein by a post-translational modification process termed SUMOylation. SUMO-1 is a highly conserved, small ubiquitin-related modifier that has been shown to be covalently conjugated to a large variety of cellular proteins (Kamitani et al., 1997; Mahajan et al., 1997; Matunis et al., 1996). Cloning of SUMO-1 was first described by Boddy et al. (1996). SUMO-2 and SUMO-3 share 95% sequence identity, but only 50% sequence identity to SUMO-1. SUMO-1 is conjugated to a target protein in a similar way to ubiquitin and has been implicated in multiple cellular processes, including nuclear transport, cell cycle control, oncogenesis, inflammation and response to viral infection. SUMO-1 conjugation forms an isopeptide bond between Gly97 at the C-terminus of SUMO-1 and the ε-amino group on the Lysine side chain of the target protein; however it is unable to form multi-chain species (Bayer et al., 1998; Mahajan et al., 1997; Mahajan et al., 1998). SUMO-1 targets substrates including RanGAP1, PML, Sp100, HSF1, Smad4, IκBα, c-Jun, p53 and Mdm2 (Melchior. 2000). RanGAP1, a Ran GTPase-activating is a major SUMO-1 substrate protein involved in nucleocytoplasmic trafficking (Swaminathan et al., 2004). SUMO-1 covalently modifies a single lysine residue at position 526 in the C-terminus of RanGAP1 (Mahajan et al., 1997; Matunis et al., 1996; Muller et al., 1998). SUMO-1 modified RanGAP1 has been found tightly associated with the nuclear envelope (Mahajan et al., 1997; Matunis et al., 1996) an observation which supports its role in nucleocytoplasmic trafficking.

References:

Bayer P, Arndt A, Metzger S, Mahajan R, Melchior F, Jaenicke R, Becker J (1998) Structure determination of the small ubiquitin-related modifier SUMO-1. J Mol Biol 280, 275-86.
Boddy MN, Howe K, Etkin LD, Solomon E, Freemont PS (1996) PIC 1, a novel ubiquitin-like protein which interacts with the PML component of a multiprotein complex that is disrupted in acute promyelocytic leukaemia. Oncogene 13, 971-82.
Kamitani T, Nguyen HP, Yeh ET (1997) Preferential modification of nuclear proteins by a novel ubiquitin-like molecule. J Biol Chem 272, 14001-4.
Mahajan R, Delphin C, Guan T, Gerace L, Melchior F (1997) A small ubiquitin-related polypeptide involved in targeting RanGAP1 to nuclear pore complex protein RanBP2. Cell 88, 97-107.
Mahajan R, Gerace L, Melchior F (1998) Molecular characterization of the SUMO-1 modification of RanGAP1 and its role in nuclear envelope association. J Cell Biol 140, 259-70.
Matunis MJ, Coutavas E, Blobel G (1996) A novel ubiquitin-like modification modulates the partitioning of the Ran-GTPase-activating protein RanGAP1 between the cytosol and the nuclear pore complex. J Cell Biol 135, 1457-70.
Melchior F (2000) SUMO–nonclassical ubiquitin. Annu Rev Cell Dev Biol 16, 591-626.