Fig. 2: Interactions between nematode serine protease inhibitor I8 (SPI-I8) with host proteins. | Communications Biology

Fig. 2: Interactions between nematode serine protease inhibitor I8 (SPI-I8) with host proteins.

From: Nematode serine protease inhibitor SPI-I8 negatively regulates host NF-κB signalling by hijacking MKRN1-mediated polyubiquitination of RACK1

Fig. 2: Interactions between nematode serine protease inhibitor I8 (SPI-I8) with host proteins.

a, b Co-immunoprecipitation analysis of Flag-fused Haemonchus contortus SPI-I8A/B with HA-fused ovine E3 ubiquitin-protein ligase makorin-1 (OaMKRN1) and/or ovine receptor of activated protein C kinase 1 (OaRACK1) in human embryo kidney (HEK) 293 T cells. c Co-immunoprecipitation analysis of Flag-fused human MKRN1 (HsMKRN1) and Myc-fused RACK1 (HsRACK1) in HEK 293 T cells. WCL, whole cell lysate; IP, immunoprecipitated protein. Beta-tubulin is used as the internal control. d Molecular docking of OaRACK1 and OaMKRN1, Hc-SPI-I8A and OaRACK1, Hc-SPI-I8B and OaRACK1, Hc-SPI-I8B and OaMKRN1, Hc-SPI-I8B, OaRACK1 and OaMKRN1 in silico. It is predicted that the binding of Hc-SPI-I8A/B and OaMKRN1 hijacks the space for OaMKRN1 to bind RACK1. Hc-SPI-I8A/B is coloured in red, ovine MKRN1 in blue, and OaRACK1 in gold. Binding free energy (ΔG) of each docking was attached. Structures of Hc-SPI-I8A/B, the ovine MKRN1 were modelled with AlphaFold2 (v. 2.1.085), and ovine RACK1 retrieved from AlphaFold Protein Structure Database. Molecular docking of molecules was performed using the ClusPro and HawkDock server. Models were displayed and superimposed using UCSF ChimeraX.

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