In this study, we report on mutational studies performed to investigate the mechanism of binding of 14-3-3 proteins to the plasma membrane H+-ATPase of plant cells. In fact, although the molecular basis of the interaction between 14-3-3 and the known mode-1 and mode-2 consensus sequences are well characterized, no information is available regarding the association with the H+-ATPase, which contains the novel binding site YTV totally unrelated to the 14-3-3 canonical motifs. To this purpose, different mutants of the maize 14-3-3 GF14-6 isoform were produced and used in interaction studies with the plasma membrane H+-ATPase and with a peptide reproducing the 14-3-3 binding site of the enzyme. The ability of 14-3-3 mutants to stimulate H+-ATPase activity was also tested. To investigate the mechanism of fusicoccin-dependent interaction, binding experiments between 14-3-3 proteins and mutants of the extreme portion of the H+-ATPase C terminus were also carried out. The results demonstrate that mutations of Lys56 and Val185 within the amphipathic groove disrupt the ability of GF14-6 to interact with H+-ATPase and to stimulate its activity. Moreover, substitution of Asp938 and Asp940 in the MHA2 H+-ATPase C terminus greatly decreased association with GF14-6, thereby demonstrating a crucial role of negatively charged residues in the fusicoccin-dependent interaction.
Visconti, S., Camoni, L., Fullone, M.R., Lalle, M., Marra, M., & Aducci, P. (2003). Mutational analysis of the interaction between 14-3-3 proteins and plant plasma membrane H+-ATPase. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 278(10), 8172-8178.
|Tipologia:||Articolo su rivista|
|Citazione:||Visconti, S., Camoni, L., Fullone, M.R., Lalle, M., Marra, M., & Aducci, P. (2003). Mutational analysis of the interaction between 14-3-3 proteins and plant plasma membrane H+-ATPase. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 278(10), 8172-8178.|
|IF:||Con Impact Factor ISI|
|Settore Scientifico Disciplinare:||Settore BIO/04|
|Revisione (peer review):||Sì, ma tipo non specificato|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1074/jbc.M211039200|
|Stato di pubblicazione:||Pubblicato|
|Data di pubblicazione:||2003|
|Titolo:||Mutational analysis of the interaction between 14-3-3 proteins and plant plasma membrane H+-ATPase|
|Autori:||Visconti, S; Camoni, L; Fullone, MR; Lalle, M; Marra, M; Aducci, P|
|Appare nelle tipologie:||01 - Articolo su rivista|