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Mechanistic dissection of increased enzymatic rate in a phase-separated compartment.
Peeples W, Rosen MK
Nat Chem Biol. 2021 Jun;17(6):693-702. doi: 10.1038/s41589-021-00801-x. Epub 2021 May 25.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
175232polyPRM5Bacterial expression and purification, mixed with polySH3 to form condensates
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175233FRB-polySH33Bacterial expression and purification, mixed with polyPRM to form condensates, can bind FKBP in the presence of rapamycin
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175234FRB-polySH35Bacterial expression and purification, mixed with polyPRM to form condensates, can bind FKBP in the presence of rapamycin
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175235EGFP-RanGAPBacterial expression and purification, high affinity SUMOylation substrate
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175236FKBP-EGFP-RanGAPBacterial expression and purification, high affinity SUMOylation substrate that can recruited to FRB with rapamycin
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175237EGFP-RanGAP*Bacterial expression and purification, low affinity SUMOylation substrate, point mutation F562A reduces affinity for E2, increasing the Km
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175238FKBP-EGFP-RanGAP*Bacterial expression and purification, low affinity SUMOylation substrate that can recruited to FRB with rapamycin, point mutation F562A reduces affinity for E2, increasing the Km
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175239ShadowG-SUMO1Bacterial expression and purification, conjugatable SUMO for SUMOylation
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175240SAE2-mGarnetBacterial expression and purification, combined with SAE1 to form the E1 enzyme heterodimer
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175241SAE1Bacterial expression and purification, combined with SAE2 to form the E1 enzyme heterodimer
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175242mCherry-E2Bacterial expression and purification, E2 enzyme (UBC9) for SUMOylation
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175243mCherry-FKBP-E2Bacterial expression and purification, E2 enzyme (UBC9) for SUMOylation, can be recruited to FRB with rapamycin, CyPet acts as a FRET donor to Ypet
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175244CyPet-FKBP-E2Bacterial expression and purification, E2 enzyme (UBC9) for SUMOylation, can be recruited to FRB with rapamycin
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175245FKBP-YPet-RanGAP*Bacterial expression and purification, SUMOylation substrate that can be recruited to FRB with rapamycin, Ypet is a FRET acceptor for CyPet
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175246EGFP-PML peptideBacterial expression and purification, low affinity SUMOylation substrate
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175247FKBP-EGFP-PML peptideBacterial expression and purification, low affinity SUMOylation substrate with a relatively high Km, can be recruited to FRB with rapamycin
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175792FRB-polySH33-EGFPBacterial expression and purification, fluorescently tagged scaffold for droplet visualization and volume quantification
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Antibodies from Article