Skip to main content
. Author manuscript; available in PMC: 2020 Dec 29.
Published in final edited form as: Nat Immunol. 2020 Jun 29;21(8):857–867. doi: 10.1038/s41590-020-0705-6

Figure 4. YopM interacts with pyrin and RSK.

Figure 4

a, b, Immunoblot analysis for pyrin (a) or YopM (b) of endogenous proteins immunoprecipitated (IP) with antibody to human RSK2 (a) or pyrin (b) from lysates (Lys) of control CD14+ monocytes with or without Y. pestis infection. c, GST-pulldown assay of purified Myc/His-tagged N-terminal (amino acids 1–330) or C-terminal (amino acids 331–781) of human pyrin with purified GST or GST-YopM. d, Schematic structure of WT YopM of Y. pestis with various mutant YopM structures used in YopM-pyrin binding and kinase assay. e, GST-pulldown assay of lysates of HEK293T cells, expressing Myc/His-tagged human pyrin, with GST-tagged WT YopM or indicated mutant YopM. f, In vitro kinase assay of purified Myc/His-tagged N-terminal human pyrin with recombinant RSK2 in the presence of GST-tagged WT YopM or indicated mutant YopM. g, In vitro kinase assay of purified Myc/His-tagged N-terminal human pyrin or PYRIN domain-deleted N-terminal pyrin (amino acids 96–330) with recombinant RSK2 in the presence of purified GST or GST-YopM, and analyzed by immunoblot with an antibody specific for phosphorylated serine. Asterisk denotes phosphorylated GST-YopM. h, Proposed model for mechanism of PKN-YopM-RSK-mediated pyrin inflammasome suppression. Data are representative of three independent experiments with similar results (a–c, e–g).