Multiple sequence alignment and phylogenetic analysis of Lpa protein.

(A) Multiple sequence alignment of Lpa1 protein from Sanggol (s-mtLpa1) and other major plant species. The P-loop kinase domain of 2-PGK found in Methanothermus fervidus is marked with a red line. P-loop is an ATP/GTP binding site motifs (Walker A and Walker B) are indicated using pink and black arrowheads, respectively. The yellow box shows a single amino acid substitution in the conserved kinase domain in lpa mutant, Sanggol. (B) Phylogenetic relationship among protein families of various kinases, including 2-PGK, serine threonine protein kinase (S/T), fructokinase (FK), myo- inositol kinase 1 (MIK), inositol 1,3,4,5,6 pentakisphosphate 2-kinase (IPK1), inositol 1,4,5-tris-phosphate kinase (IPK2), hexokinase 1 (HXK), and hexokinase 2, with the Lpa protein clade. Phylogenetic tree was constructed using amino acid sequences of Lpa and other kinase protein families from selected species using the neighbour-joining method with 1,000 bootstrap replicates. Protein homologs are indicated with an ‘H’. ob, Oryza brachyantha; Os-J, Oryza sativa L. japonica; Osi, Oryza sativa L. indica; Agt/Ast, Aegilops tauschii; sb, Sorghum bicolor; zm, Zea Mays; Si, Setaria italica; Ao, Asparagus officinalis; Ga, Gossypium arboretum; Cc, Cajanus cajan; Gm, Glycine max; Cm, Cucurbita maxima; ta, Nicotiana tabacum; Bn, Brassica napus; At, Arabidopsis thaliana; Ac, Ananas cosmos; Ztm, Zotera marina; Ci, Chrysanthemum indicum; Mf, methanothermus fervidus; R-570, Saccharum; Al, Arabidopsis lyrata.