A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins
Articolo
Data di Pubblicazione:
2002
Citazione:
A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins / P. Morandini, M. Valera, C. Albumi, M.C. Bonza, S. Giacometti, G. Ravera, I. Murgia, C. Soave, M.I. De Michelis. - In: PLANT JOURNAL. - ISSN 0960-7412. - 31:4(2002), pp. 487-497. [10.1046/j.1365-313X.2002.01373.x]
Abstract:
Using the two-hybrid technique we identi®ed a novel protein whose N-terminal 88 amino acids (aa)
interact with the C-terminal regulatory domain of the plasma membrane (PM) H+-ATPase from
Arabidopsis thaliana (aa 847±949 of isoform AHA1). The corresponding gene has been named Ppi1 for
Proton pump interactor 1. The encoded protein is 612 aa long and rich in charged and polar residues,
except for the extreme C-terminus, where it presents a hydrophobic stretch of 24 aa. Several genes in
the A. thaliana genome and many ESTs from different plant species share signi®cant similarity (50±70%
at the aa level over stretches of 200±600 aa) to Ppi1. The PPI1 N-terminus, expressed in bacteria as a
fusion protein with either GST or a His-tag, binds the PM H+-ATPase in overlay experiments. The same
fusion proteins and the entire coding region fused to GST stimulate H+-ATPase activity. The effect of the
His-tagged peptide is synergistic with that of fusicoccin (FC) and of tryptic removal of a C-terminal
10 kDa fragment. The His-tagged peptide binds also the trypsinised H+-ATPase. Altogether these results
indicate that PPI1 N-terminus is able to modulate the PM H+-ATPase activity by binding to a site
different from the 14-3-3 binding site and is located upstream of the trypsin cleavage site.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
plasma membrane H+-ATPase ; fusicoccin ; Arabidopsis thaliana ; two-hybrid ; 14-3-3 protein ; fusion protein.
Elenco autori:
P. Morandini, M. Valera, C. Albumi, M.C. Bonza, S. Giacometti, G. Ravera, I. Murgia, C. Soave, M.I. De Michelis
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