K-712

Material Data Sheet

SUMO1 Mutant Protein Set

Price: $395.00
Catalog #: K-712

SUMO1 Mutant Protein Set

Price: 
$395
Quantity: 
1 Kit

The ubiquitin-like SUMO-1, SUMO-2 and SUMO3 proteins are conjugated to a variety of proteins in the presence of UbcH9 and the SAE1/SAE2 (human) or Aos1/Uba2 (yeast) activating enzyme. SUMO modification has been implicated in diverse functions such as nuclear transport, chromosome segregation and transcriptional regulation, apoptosis and protein stability. Human SUMO-1 shares 46% and 47% identity with SUMO-2 and SUMO-3 respectively and does not contain the exact ψΚXE consensus sequence found in SUMO-2 and SUMO-3. Within this sequence ψ represents a large hydrophobic amino acid (I, L, or V), K is the lysine that becomes modified, X is any residue and E is glutamic acid. Many known SUMO-1 conjugation sites occur within this motif, but SUMOylation also occurs on lysine residues located within non-consensus regions. SUMO-1 has been shown to form chains in vitro and in vivo but often the linkage is uncharacterized, and the function of SUMO chains has not yet been fully elucidated. SUMO-1 multimerization in vitro has been shown to occur predominantly via K7, K16 and K17. The proteins in this kit have these lysine residues mutated to arginine and can be used to investigate mono-SUMOylation requirements or to reduce poly-SUMO chain formation. Included is wild-type SUMO-1 to be used as a positive control in SUMO-1 conjugation assays which can be performed using the SUMO-1 Conjugation Kit (K-710).

Product Information

Stock: 
50mM HEPES pH 8.0, 150mM NaCl, 1 mM DTT.
Purity: 
> 95 % by SDS-PAGE
Supplied: 
ProteinMWConcentrationQuantitySUMO-1 wildtype11.1 kDaX mg/ml (X μM)50 μgSUMO-1 K7R11.1 kDaX mg/ml (X μM)50 μgSUMO-1 K16R11.1 kDaX mg/ml (X μM)50 μgSUMO-1 K17R11.1 kDa X mg/ml (X μM)50 μgSUMO-1 K7R K16R11.1 kDaX mg/ml (X μM)50 μg SUMO-1 K7R K17R11.1 kDaX mg/ml (X μM) 50 μg SUMO-1 K16R K17R11.1 kDaX mg/ml (X μM) 50 μgSUMO-1 K7R K16R K17R11.1 kDaX mg/ml (X μM)50 μg
Background: 
The ubiquitin-like SUMO-1 is conjugated to a variety of proteins in the presence of UbcH9 and the SUMO E1 activating enzyme (SAE1/SAE2 in human, or Aos1/Uba2p in yeast). The heterodimeric SAE1/SAE2 complex (38 and 70 kDa respectively) uses ATP to adenylate the C-terminal glycine<br>residue of SUMO-1, forming a high-energy thiolester bond with the SAE2 subunit. The second step is the trans-esterification reaction whereby the activated SUMO-1 is transferred to Cys93 of UbcH9. UbcH9 is a member of the E2 family and is homologous to ubiquitin conjugating enzymes, but is<br>specific for the conjugation of SUMO to a variety of target proteins. This E2 is unusual in that it interacts directly with protein substrates that are modified by sumolyation, and may play a role in substrate recognition. Sumoylated substrates are primarily localized to thenucleus (RanGAP-1, RANBP2, PML, p53, Sp100, HIPK2) but there are also cytosolic substrates (IκBα, GLUT1, GLUT4). SUMO modification has been implicated in functions such as nuclear transport, chromosome segregation and transcriptional regulation, apoptosis and protein function and stability.

Use & Storage

Use: 

Typical concentration to support conjugation reaction in vitro is 10 μM-50 μM depending on conditions. 

Storage: 

Store at -80°C. Avoid multiple freeze/thaw cycles.

Literature

References: 

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