Specifications & Handling:

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Small Ubiquitin-related Modifier 1; GAP-modifying Protein 1; SMT3C, SMT3H3, UBL1; Sentrin; Ubiquitin-homology Domain Protein PIC1

Human

20µl of 10X SUMO E1 Enzyme20µl of 10X SUMO120µl of 10X Ubc9 (UBE2I)20µl of 10X Reaction Buffer20µl of 10X Mg-ATP SolutionNOTE: Kit contains reagents sufficient for 5-10 x 20μl reactions.

Use: This kit is designed for the conjugation of the ubiquitin-like modifier SUMO-1 to protein substrates in vitro, which requires the activities of the heterodimeric human E1 activating enzyme (SAE1/SAE2) and the Ubc9 E2 enzyme.

Manufactured by Boston Biochem

Kit

DRY ICE

-20°C

-80°C

Aliquot to avoid freeze/thaw cycles.

Stable for at least 1 year after receipt when stored at -80°C.

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Description:

The ubiquitin-like SUMO-1 is conjugated to a variety of proteins in the presence of Ubc9 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 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 Ubc9. Ubc9 is a member of the E2 family and is homologous to ubiquitin conjugating enzymes, but is 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 the nucleus (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, protein function and stability.

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