A number of putative phosphorylation sites at regulatory C-domain of H+-ATPase (AHA2) have been point-mutated to alanine residues (to prevent possible phosphorylation) or aspartate residues (to mimic phosphorylation of residue) and the mutated aha2 enzyme expressed in the yeast strain RS-72. Most of the mutations show positive or negative effect on yeast growth in functional complementation assays. It shows in vivo functioning of the residues and suggests, that plant H+-ATPase could be regulated by phosphorylation at several sites being in yeast cells.Plant H+-ATPase purified from yeast cells by his-tag affinity chromatography was subjected to IMAC and TiO2 for enrichment of phosphopeptides. The phosphopeptides were analyzed by LC/MS/MS. Phosphorylation sites identified in vivo in the heterologous host are compared to the functional analysis of the aha2 point mutations.
|Status||Udgivet - 2008|
|Begivenhed||12th International ATPase Conference - Aarhus, Danmark|
Varighed: 5. aug. 2008 → 10. aug. 2008
|Konference||12th International ATPase Conference|
|Periode||05/08/2008 → 10/08/2008|