Fig. 1 - Supplemental 3

Optimization of Halo-PafA. (A) Schematic illustration of Halo-PafA derivatives: Halo8KR-PafA, which includes eight lysine-to-arginine mutations to abolish self-pupylation, and Halo-PafAS126A, which contains a serine-to-alanine mutation at position 126 to decrease depupylase activity of PafA. (B) In vitro self-pupylation results show that Halo8KR-PafA exhibits minimal pupylation levels, similar to PafA alone. Halo-PafAS126A demonstrates reduced self- and polypupylation while enhancing multipupylation. Reactions were conducted with 1 μM of a Halo-PafA derivative and 10 μM of TS-sPup or TS-sPupK61R at 37°C for 30 min. Yellow arrowheads indicate polypupylated bands, while black arrowheads and arrows indicate multipupylated bands and Halo-PafA, respectively.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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