Table 3.
CaM-binding site prediction comparison for the region homologous to B. distachyon amino acids 334–351
Sequences were obtained from the NCBI Database and aligned using Clustal Omega (54). Regions homologous to the predicted BdALMT12 CBD were evaluated using the calmodulation database and Meta-analysis Predictor software to analyze the full protein sequence (35). The higher the positional prediction score numbers, the higher the likelihood for a binding site, with scoring letters being a continuation of the numbers with A = 10, B = 11, C = 12, etc. Amphipathicity of the helices is expressed as the absolute value of the 3D hydrophobic moment of the helix, calculated using the 3D-HM prediction software, and is expressed as Å·kT/e, where kT/e is the electrostatic potential, and Å is the length of the vector (49).
| Species | Annotation | NCBI accession no. | Motif starting residue no. | Putative CBD motif | CaM motif positional prediction scores | Hydrophobic moment vector (Å·kT/e) |
|---|---|---|---|---|---|---|
| B. distachyon | BdALMT12 | XP_003574370 | 334 | TRVAREVAKVLQELAVSI | 55A999CAAA98898946 | 12.479 |
| Z. mays | ZmALMT12 | XP_008648985 | 338 | TRVAREVAKVLQELAASI | 557666755543343335 | 12.512 |
| O. sativa | OsALMT12 | XP_015614622 | 334 | TRVAREVVKVLQELAVSI | 66BAAADBAA98898946 | 12.405 |
| S. bicolor | SbALMT12 | XP_002437417 | 337 | TRVAREVAKVLQELAVSI | 55A999CAAA98898946 | 12.479 |
| A. tauschii | AgtALMT12 | XP_020161007 | 334 | TRVAREVTKVLQELADSI | 557666755543343335 | 10.084 |
| A. comosus | AcALMT12 | OAY77120 | 309 | IRVAGEVSKVLIELARSI | A8CBBBCAAA87453335 | 8.570 |
| A. officinalis | AoALMT12 | XP_020261302 | 325 | TRVATEVSKVLLELGTSI | 559888977787565557 | 8.206 |
| V. vinifera | AvALMT12 | XP_002278594 | 345 | FQVAAEVSKVLKELANCI | B8A999A88854453335 | 7.002 |
| S. oleracea | SoALMT12 | XP_021854304 | 340 | IRLAAEVSKVVRELANSI | B8A999A88854453335 | 7.840 |
| N. tabacum | NtALMT12 | XP_016493112 | 340 | IRLAGEVTKALKELGDSI | 857666755554453335 | 8.815 |
| G. max | GmALMT12 | XP_003516635 | 344 | IRLAAEVSKVLIELSNSI | B8CBBBCAAA87453335 | 6.646 |
| S. lycopersicum | SlALMT12 | XP_004244701 | 339 | IRLSGEVAKALKELGDSI | 968777866665563335 | 8.329 |
| A. thaliana | AtQUAC1 | O49696 | 331 | VRLAGEVCKALTELADSI | 857666755554453335 | 8.872 |
| T. aestivum | AtALMT1 | Q76LB1 | 324 | GEMSLHSSKVLRDLAMAT | 333342222554476666 | 4.941 |