The C-terminal sequence from common bugle leaf galactan:galactan galactosyltransferase is a non-sequence-specific vacuolar sorting determinant

The Ajuga reptans L. galactan:galactan galactosyltransferase (ArGGT) is a vacuolar enzyme that synthesizes long-chain raffinose family oligosaccharides (RFOs), the major storage carbohydrates of this plant. ArGGT is structurally and functionally related to acid plant alpha-galactosidases (alpha-Gals) of the glycosylhydrolase family 27, present in the apoplast or the vacuole. Sequence comparison of acid alpha-Gals with ArGGT revealed that they all contain an N-terminal signal sequence and a highly similar core sequence. Additionally, ArGGT and some acid alpha-Gals contain C-terminal extensions with low sequence similarities to each other. Here, we show that the C-terminal pentapeptide, SLQMS, is a non-sequence-specific vacuolar sorting determinant. Analogously, we demonstrate that the C-terminal extensions of selected acid alpha-Gals from Arabidopsis, barley, and rice, are also non-sequence-specific vacuolar sorting determinants, suggesting the presence of at least one vacuolar form of acid alpha-Gal in every plant species.