Phospholipase cation requirement

Cation Requirements. While some of the phospholipases C found in bacteria appear to prefer Ca2+, there are many many reports supporting Zn2+ as the divalent cation of choice. There is some support for the fact that this enzyme is probably a metallo (Zn2+) protein which also requires Ca2+ for catalytic activity, but there is more evidence for the enzyme s ability to influence the surface charge on the micellar substrate system. 

Both intracellular release and transmembrane flux contribute to the rise in intracellular Ca2+.14,15 The rise in keratinocyte intracellular Ca2+ in response to raised extracellular Ca2+ has two phases (a) an initial peak, not dependent on extracellular Ca2+ and (b) a later phase that requires extracellular Ca2+.14 An early response of human keratinocytes to increases in extracellular Ca2+ is an acute increase in intracellular Ca2+. Stepwise addition of extracellular Ca2+ to neonatal human keratinocytes is followed by a progressive increase in intracellular Ca2+, where the initial spike of increased intracellular Ca2+ is followed by a prolonged plateau of higher intracellular Ca2+.16 The response of intracellular Ca2+ to increased extracellular Ca2+ in keratinocytes is saturated at 2.0 mM extracellular Ca2+.16,17 The response of intracellular Ca2+ to increased extracellular Ca2+ in keratinocytes resembles the response in parathyroid cells, in that a rapid and transient increase in intracellular Ca2+ is followed by a sustained increase in intracellular Ca2+ above basal level. This multiphasic response is attributed to an initial release of Ca2+ from intracellular stores followed by an increased influx of Ca2+ through voltage-independent cation channels. The keratinocyte and parathyroid cell contains a similar cell membrane calcium receptor thought to mediate this response to extracellular Ca2+. This receptor can activate the phospholipase-C pathway, leading to an increase... 

Like the lipase, the phospholipase hydrolyzes its substrates at an interface in this case the interface of micelles and water. However pure phospholipid micelles are not digested, and even micelles in which the substrate molecules are spaced apart by inclusion of solvents such as ether or by cationic detergents are not attacked. Micelles containing anionic detergents such as bile salts are attacked. The enzyme requires a negative surface charge on the micelle even though the substrate molecule itself may be an electrically neutral lipid such as phosphatidyl choline. 

In higher plants, the lipolytic enzymes and their physiological functions are not well characterized [1]. iMost reports demonstrated that phospholipid catabolism in plants is achieved by the concerted actions of membrane-bound enzymes including phospholipase D, phosphatidate phosphatase, lipolytic acyl hydrolases and lipoxygenases [1,2]. With the exception of the phospholipase D, the literature on plant phospholipases is still very limited. We previously reported that tonoplast from Acer pseudoplatanus cells contains small amounts of phosphatidic acid and lysophospholipids, which were produced together with free fatty acids, particularly after addition of Ca " [31. These data suggested the possible involvement of phospholipase D and phospholipase A in the metabolism of vacuolar membrane lipids. The phospholipase activities were studied by following the hydrolysis of added sn-2-[14c]linoleyl-PC to tonoplast vesicles. Tonoplast was obtained by osmotic lysis of pure preparations of vacuoles isolated from protoplasts derived from Acer pseudoplatanus cells [4]. This present work demonstrated clearly the presence of phospholipase D and phospholipase Ai activities associated with the tonoplast of Acer, The phospholipase Ai showed an optimal activity at pH about 6-6.5, did not necessarily require divalent cations, but was stimulated by Mg- and particularly by Ca. This work presents the first evidence for the presence of phospholipases A in plant cells. 

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