Leptospira Sph2 Epitope Prediction and Analysis

To predict B-cell linear epitopes and perform in silico analyses, we used the entire sequence of Sph2 of L. interrogans serogroup Icterohaemorrhagiae and serovar Lai (Uniprot ID: P59116), obtained from the Uniprot database https://www.uniprot.org/ (accessed on 24 March 2020), as the sequence reference to this study. Moreover, to evaluate the similarity of Sph2 among pathogenic leptospires, we compared the used sequence with the described sequences of Sph2 from L. interrogans (serogroups: Australis, Bataviae, and Pyrogenes); L. alexanderi (serogroup: Manhao); L. alstonii (serogroup: Ranarum); L. borgpetersenii (serogroups: Pomona and Sejroe); L. noguchii (serogroups: Autumnalis and Panama); L. kirschneri (serogroup: Autumnalis); L. santarosai (serogroup: Javanica); and L. weilii (serogroups: Ranarum and Tarassovi). In addition, to investigate the conservation of reference Sph2 and other Leptospira spp. sphingomyelinases, this sequence was also aligned and compared with sequences of SphH (Uniprot ID: O34095), Sph1 (Uniprot ID: P59115), Sph3 (Uniprot ID: A0A0E2DC81), and Sph4 (Uniprot ID: A0A0E2DCF7) from L. interrogans serogroup Icterohaemorrhagiae and serovar Lai. Finally, the Sph2 reference sequence was also compared with sphingomyelinases from Listeria ivanovii (Uniprot ID: Q9RLV9), Bacillus cereus (Uniprot ID: P09599), Staphylococcus aureus (Uniprot ID: A0A7U4AUV1), and Pseudomonas sp. (Uniprot ID: Q93HR5). All studied sequences were accessed on 24 March 2020 and are summarized in Table 1.