Homology search for MmSPO16 orthologs and related proteins was performed using PSI-BLAST and HHpred (http://toolkit.tuebingen.mpg.de/). Alignment of SPO16 orthologs were performed with Clustal-Omega (www.ebi.ac.uk/Tools/msa/clustalo/) and colored with ESPript (http://espript.ibcp.fr/) using the Risler homology matrix (with a global score of 0.5). Secondary structure prediction was performed using the PSIPRED (http://bioinf.cs.ucl.ac.uk/psipred/), while homology modeling was performed with I-TASSER (https://zhanglab.ccmb.med.umich.edu/I-TASSER/) (3537). The C-score for the secondary structure of MmSPO16 is −1.67. Protein structure comparison was performed with FATCAT (http://fatcat.burnham.org/). The structure alignment for MmSPO16 and ApXPF has 177 equivalent positions with a root mean square deviation (RMSD) of 2.35, without twists (P = 2.22 × 10–16). The structure alignment for MmSPO16 and ScSPO16 has 126 equivalent positions with an RMSD of 3.07, with four twists (P = 0.0264).

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