Phospholipid Requirement of the Vanadate-Sensitive ATPase from Maize Roots Evaluated by Two Methods

Abstract

The activation of the vanadate-sensitive ATPase from maize (Zea mays L.) root microsomes by phospholipids was assessed by two different methods. First, the vanadate-sensitive ATPase was partially purified and substantially delipidated by treating microsomes with 0.6% deoxycholate (DOC) at a protein concentration of 1 milligram per milliliter. Vanadate-sensitive ATP hydrolysis by the DOC-extracted microsomes was stimulated up to 100% by the addition of asolectin. Of the individual phospholipids tested, phosphatidylserine and phosphafidylglycerol stimulated activity as much as asolectin, whereas phosphatidylcholine did not. Second, phospholipid dependence of the ATPase was also assessed by reconstituting the enzyme into proteoliposomes of differing phospholipid composition. In these experiments, the rate of proton transport and ATP hydrolysis was only slightly affected by phospholipid composition. DOC-extracted microsomes reconstituted with dioleoylphosphatidylcholine had rates of proton transport similar to those found with microsomes reconstituted with asolectin. The difference between the two types of assays is discussed in terms of factors contributing to the interaction between proteins and lipids.

Lysophosphatidylcholine specifically stimulates plasma membrane H+‐ATPase from corn roots

“…tergent [3][4][5][6][7][8][9][10][11][12][13][14] Detergent used for solubilization ATPane activity (.amol I"l!llS I,I". 'I'I'I'3,.N No+c~ld+cr 1~,~1 exceeded that obtained by soybean phospholipids (not shown), We have investigated the influence of lysoPC on some properties of H+-ATPase, The maximal stimulation by lysoPC occurred at pH 6,5, which ¢oin¢ided with the pH optimum of the enzyme, but the effect reversed to inhibitory at pH > 7,0 (Fig, 1), LysoPC also did not change tile sensitivity of tile enzyme to its specific inhibitor vanadate, We also found that lysoPC had no effect on the acid phosphatase activity of plasma membrane (Table III).…”

“…When considering different ways of regulation, the effects of lipids on this integral enzyme are of special interest. The lipid dependence of the plasma membrane H +-ATPase activity has been previously demonstrated, both on partially delipidated [3] and solubilized preparations [4,5]. The enzyme reactivation by the different phospholipids revealed that the lysoPC was the most powerful activator in plasma membrane preparations of plants and yeast [4,6].…”

Mechanism for the Activation of Plasma Membrane H+-ATPase from Rice (Oryza sativa L.) Culture Cells by Molecular Species of a Phospholipid

“…30-35%) in the plasma membrane (10,14) and activated ATPase activity (Table I) (10). Although previous studies have examined the requirement of phospholipids to induce the ATPase activity using delipidated membrane vesicles (2), in this study we have completely solubilized the ATPase with a two-step procedure using deoxycholate and zwittergent prior to lipid additions. Serrano et al (18) performed a less extensive examination of the lipid requirements of a cholate-extracted preparation of the plant plasma membrane H+-ATPase and the protein content of this preparation was unclear.…”

“…The mechanism for regulation of the ATPase by auxin is unknown (17). In our previous paper (10) and other reports (2,3,18), plasma membrane H+-ATPase was shown to require phospholipids for its activation, indicating that its activity may be regulated by interactions between this protein and the phospholipid environment in the plasma membrane. However, the effects…”

Plant lipid environment and membrane enzymes: the case of the plasma membrane H+-ATPase

“…No effect Arabidopsis thaliana Gomès et al (1996) Spinacia olerancea Johansson et al (1994) Glycerolipids Asolectin Stimulation Oryza sativa Kasamo (1990) Spinacia olerancea Johansson et al (1994) Vigna radiata Kasamo and Nouchi (1987), Kasamo and Yamanishi (1991) Zea mays Brauer and Tu (1989) Phosphatidylcholine (PC) Stimulation Avena sativa Serrano et al (1988) Oryza sativa Kasamo (1990) Vigna radiata Kasamo and Nouchi (1987), Kasamo and Yamanishi (1991) Phosphatidylglycerol (PG) Stimulation Avena sativa Serrano et al (1988) Oryza sativa Kasamo (1990) Raphanus sativus Coccuci and Marré (1984) Vigna radiata Kasamo and Nouchi (1987), Kasamo and Yamanishi (1991) Zea mays Brauer and Tu (1989) Phosphatidylserine (PS) Stimulation Avena sativa Serrano et al (1988) Oryza sativa Kasamo (1990) Raphanus sativus Coccuci and Marré (1984) Vigna radiata Kasamo and Nouchi (1987), Kasamo and Yamanishi (1991) Zea mays Brauer and Tu (1989) Phosphatidic acid (PA) Stimulation Avena sativa Serrano et al (1988) Zea mays Brauer and Tu (1989) Inhibition Oryza sativa Kasamo (1990) Vigna radiata. Kasamo and Nouchi (1987), Kasamo and Yamanishi (1991) Plant Cell Rep…”