Fig. 2: HmwC affects global O-GlcNAcylation in Y. pestis.
From: A protein O-GlcNAc glycosyltransferase regulates the antioxidative response in Yersinia pestis
a O-GlcNAcylation of proteins in ΔhmwC was significantly lower than that in WT. The experiment was independently repeated at least three times, yielding consistent results. Equal protein samples obtained from the wild-type Y. pestis and ΔhmwC grown under different conditions were separated by 12% SDS-PAGE and immunoblotted with a mouse anti-O-GlcNAc antibody to detect O-GlcNAcylated proteins. b Protein structure prediction and domain division of HmwC by AlphaFold2. Structural domains were divided according to protein annotations and represented with different colors. c The secondary structure of HmwC based on PDB:3Q3E (https://doi.org/10.2210/pdb3Q3E/pdb) template alignment using Espript 3.0 online tool. Identical and conserved residues were highlighted in red background and light red font. The helices and folds observed in the structures were indicated above the sequences. d Phylogenetic tree of HmwC homologous sequences in bacteria. The homologous sequences of HmwC in different species were retrieved using BLAST at NCBI. Sequences with more than 40% identity were selected for phylogenetic tree analysis. HmwC is represented by Y. pestis in group I and highlighted in red font. ApHMW1C by A. pleuropneumoniae and HMW1C by H. influenzae in group III are highlighted in purple font. The evolutionary history was inferred using the Neighbor-Joining method. The bootstrap consensus tree inferred from 1000 replicates was taken to represent the evolutionary history of the taxa analyzed. The evolutionary distances were computed using the p-distance method and are in the units of the number of amino acid differences per site. Evolutionary analyses were conducted in MEGA11. The phylogenetic tree was beautified using the iTOL online tool. e Proteins in Group I and HMW1C in H. influenzae and ApHMW1C in A. pleuropneumoniae (Group III) were aligned with Uniprot, and the key sites were highlighted with a black box. Thr-438 of ApHMW1C and Ser-435 of HmwC were labeled in the tertiary structure of the protein.
