Modulation of plant plasma membrane H+-ATPase by phytotoxic lipodepsipeptides produced by the plant pathogen Pseudomonas fuscovaginae.
Pseudomonas fuscovaginae produces the lipodepsipeptides syringotoxin, fuscopeptin A and fuscopeptin B concurrently. These phytotoxins inhibit acidification of the external medium by fusicoccin-treated rice leaf sheath discs. When tested in vitro on H+-ATPase of rice shoot plasma membranes, syringotoxin and its structural analogue syringomycin, produced by P. syringae pv. syringae, displayed a double effect. At low concentrations they stimulated the ATPase activity of native right-side-out membrane vesicles in a detergent-like manner. At higher concentrations, however, this stimulation was reversed. With membranes treated with the detergent Brij 58, inhibition of ATPase activity was observed at low concentrations of the nonapeptides. The latter effect required the presence of an intact lactone ring formed by the nonapeptide head of these molecules. In contrast, fuscopeptins A and B inhibited enzyme activity regardless of the orientation of the vesicles. These observations were confirmed using plasma membranes from a yeast strain whose own H+-ATPase had been replaced by a single plant H+-ATPase isoform, PMA2, from Nicotiana plumbaginifolia. The kinetics of inhibition induced by the most active compound fuscopeptin B, showed a non-competitive pattern, with a Ki of about 1 microM. The combination of syringotoxin (or syringomycin) with the more hydrophobic fuscopeptins, in amounts with little or no effect, resulted in strong inhibition of the enzyme activity of rice membranes, suggesting a synergistic effect for the two types of toxins.
Laboratoire de Cytogénétique, Place Croix du Sud 5, B-1348 Louvain-la-Neuve, Belgium.
H
H
Batoko
A
A
de Kerchove d'Exaerde
J M
JM
Kinet
J
J
Bouharmont
R A
RA
Gage
H
H
Maraite
M
M
Boutry
D000595:Amino Acid Sequence; D001427:Bacterial Toxins; D002462:Cell Membrane / Q000201:enzymology*; D002592:Cetomacrogol / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006027:Glycosides / Q000494:pharmacology; D007700:Kinetics; D015394:Molecular Structure; D012275:Oryza / Q000201:enzymology / Q000648:ultrastructure; D010456:Peptides, Cyclic / Q000737:chemistry / Q000494:pharmacology*; D010944:Plants / Q000201:enzymology / Q000648:ultrastructure*; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011549:Pseudomonas / Q000378:metabolism
D001427:Bacterial Toxins; D004791:Enzyme Inhibitors; D006027:Glycosides; D010456:Peptides, Cyclic; C098810:fuscopeptin A; C098811:fuscopeptin B; C007808:fusicoccin; C069036:syringotoxin B; D002592:Cetomacrogol; D006180:Proton-Translocating ATPases