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Do Plants Have Ouabain (OU)-Sensitive ATPases?

In the 1960s and 1970s, many plant physiologists labored under the assumption that any electrogenic ion pumps discovered in plant cells would be similar, if not identical, to the electrogenic Na, K-ATPase found in the plasma membrane of animal cells. These expectations eventually subsided as evidence, now incontrovertible, mounted that electrogenesis in plants involves primarily OU-insensitive H-ATPases. Before this consensus was reached, however, many biochemical and physiological studies were performed concerning the effects of OU on plants. Although these results were not entirely negative, interest in the question of the mode of action of OU on plants came to a virtual standstill as researchers focused their attentions on elucidating the properties of the dominant OU-insensitive HATPases. It is not the purpose of this review to dispute the fundamental importance of OU-insensitive H ATPases in plant function, but rather to suggest that there may be some minor ATPases within higher plants that possess sensitivity to OU. A survey of the botanical literature has found more than 50 published reports consistent with the idea that pharmacologically valid concentrations of OU (i.e. 125 m or less) have physiological effects on plants or inhibit plant membrane ATPase activities in vitro. It is regrettable, therefore, that studies of the effects of OU on plants virtually ceased in the 1990s, even though there is now a whole new arsenal of molecular techniques that could be brought to bear on the question of the sites of OU action in plants. It is hoped that this review will revitalize interest in this moribund question. ATPase, the gastric H, K-ATPase, and the sarcoplasmic reticulum Ca2ATPase) are all related members of a larger molecular familythe P-type ATPases (Green, 1992). The fact that at least one other P-type ATPase besides the Na, K-ATPase exhibits sensitivity to OU, namely the H, K-ATPase of colonocytes (Cougnon et al., 1996), raises the possibility that the OUsensitive ATPases in plants need not necessarily act as Na pumps. Experiments have shown that there may be less distinction between the various types of P-type ATPases than their essentialist names would suggest. For example, the Na, K-ATPase of animal cells can, under certain conditions, pass protons instead of Na ions (Polvani and Blostein, 1988), and, conversely, the gastric H, K-ATPase can, under certain conditions, pass Na ions instead of protons (Polvani et al., 1989). Thus, researchers should be attentive to the possibility that OU-sensitive ATPases in plants could be serving slightly or very different functions from their counterparts in animal cells. Reports that OU affects physiological processes in ciliates (Fabczak and Fabczak, 1989), euglenozoa (Mikolajczyk, 1986), fungi (Ayres et al., 1987; Rauforova et al., 1997), green algae (Meszes and Erdei, 1969), and cyanobacteria (Iwano, 1995) suggest that OU sensitivity may have had an ancient origin during the molecular phylogeny of P-type ATPases. root pressure (Mozhaeva et al., 1979), transpiration (Morant-Avice et al., 1997), and flowering (Oota, 1974).

Two OU-Sensitive ATPases?

Two studies (Brown et al., 1964; Thomas, 1970) produced OU dose response curves very similar to those commonly found in animal studies. In other cases (e.g. Nassery and Baker, 1972), the dose response curves for OU sensitivity are shifted over more than 3 orders of magnitude. This suggests that there are may be two OU-sensitive processes in plants: one (type 1) with an I50 of approximately 0.1 m, and the other (type 2) with a concentration for half-inhibition (I50) of approximately 100 m. Ootas (1974) finding of an unusual U-shaped dose response curve suggests that the Lemna gibba flowering response he studied may be lowered by type 1 OU inhibition and enhanced by type 2 OU inhibition. Other evidence that there may be more than one site of OU sensitivity in plants comes from biochemical studies. Lindberg (1982) concluded that two forms of OU-sensitive ATPases exist in sugar beet (Beta vulgaris) roots; one with an optimal pH of 5.5 to 6.0, and the other with an optimal pH of 8.0. It has been hypothesized that the acidophilic OU-sensitive ATPase is endomembranous. It may be germane in this regard that plants do have a P-type Ca2-ATPase coded for by a single gene, which is found in both the plasma membrane and the vacuolar membrane (Ferrol and Bennett, 1996). Because OU interacts with the extracellular face of the Na, K-ATPase, researchers have tended to ignore the effects of intracellular OU. OU, however, is also taken up internally into animal cells (Nunez-Duran et al., 1988). The possibility that OU may have an intracellular site of action in plants is one that should be considered. LITERATURE CITED Ayres R, Steinkraus KH, Olek A, Farr D (1987) Characterization of the semi-purified membrane-bound-

Physiological Effects of OU
OU has been found to influence many physiological processes and parameters in plants, including membrane potential (Stolarek, 1968a; Latala, 1975; Zientara, 1983; Solic and Zuvic, 1990), Na fluxes (Cram, 1968; Ozerskii, 1969; Davis and Jaworski, 1979), K fluxes (MacRobbie, 1962; Stolarek, 1968b), amino acid uptake (Basu et al., 1988), electrical signaling (Desbiez, 1973; Pazurkiewicz-Kocot and Stolarek, 1980; Karege et al., 1982; Slesak, 1984), stomatal aperture (Thomas, 1970; Turner, 1973), pulvinar function (Watanabe, 1971; Jonas, 1976) respiration (Legros et al., 1973; Migliaccio 1973) secretion (Brown et al., 1964),

Origins of OU Sensitivity
The H-ATPases of plants and fungi, the Na, K-ATPase of animals, as well as certain other types of ATPases (e.g. the plasma membrane Ca 2 www.plantphysiol.org/cgi/doi/ 10.1104/pp.900046. 4

Plant Physiology, September 2002, Vol. 130, pp. 45, www.plantphysiol.org 2002 American Society of Plant Biologists

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Peter V. Minorsky Department of Natural Sciences Mercy College Dobbs Ferry, NY 10522
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