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Literature summary for 3.4.22.68 extracted from

  • Kuo, D.; Nie, M.; De Hoff, P.; Chambers, M.; Phillips, M.; Hirsch, A.M.; Courey, A.J.
    A SUMO-Groucho Q domain fusion protein: characterization and in vivo Ulp1-mediated cleavage (2011), Protein Expr. Purif., 76, 65-71.
    View publication on PubMedView publication on EuropePMC
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Application

Application Comment Organism
molecular biology modification of recombinant proteins by SUMOylation often dramatically increases solubility and stability during expression of the fusion proteins in bacteria relative to unfused proteins. After expressing a protein as a fusion to SUMO, it is often desirable to cleave the SUMO off of the fusion protein using a SUMO-specific protease such as Ulp1. To facilitate such processing, a dual expression vector is constructed encoding two fusion proteins: one consisting of SUMO fused to Ulp1 and a second consisting of SUMO fused to a His-tagged protein of interest. The SUMO-Ulp1 cleaves both itself and the other SUMO fusion protein in the bacterial cells prior to lysis, and the proteins retain solubility after cleavage, method evaluation, overview Drosophila melanogaster

Cloned(Commentary)

Cloned (Comment) Organism
expression of SUMO-S-GroQ fusion protein, i.e. SUMO fused to the Q domain of Drosophila melanogaster Groucho, containing sequences encoding the mature form of SUMO followed by a 6-His tag and a multiple cloning site harbouring the fused protein S-GroQ, co-expression of SUMO-fused SUMO-specific protease Ulp1 Drosophila melanogaster

Protein Variants

Protein Variants Comment Organism
additional information modification of recombinant proteins by SUMOylation often dramatically increases solubility and stability during expression of the fusion proteins in bacteria relative to unfused proteins. After expressing a protein as a fusion to SUMO, it is often desirable to cleave the SUMO off of the fusion protein using a SUMO-specific protease such as Ulp1. To facilitate such processing, a dual expression vector is constructed encoding two fusion proteins: one consisting of SUMO fused to Ulp1 and a second consisting of SUMO fused to a His-tagged protein of interest. The SUMO–Ulp1 cleaves both itself and the other SUMO fusion protein in the bacterial cells prior to lysis, and the proteins retain solubility after cleavage, method evaluation, overview Drosophila melanogaster

Organism

Organism UniProt Comment Textmining
Drosophila melanogaster
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged SUMO-S-GroQ by nickel affinity chromatography to over 95% purity, SUMO is clevaed off by co-expressed SUMO-fused SUMO-specific protease Ulp1 Drosophila melanogaster

Synonyms

Synonyms Comment Organism
SUMO-specific protease
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 Drosophila melanogaster
Ulp1
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 Drosophila melanogaster