Ion-Transporting ATPases

Sarcoplasmic reticulum Ca2+-transporting ATPase (SR-ATPase) is a member of the ATPase family that couples ion translocation across cell membranes to hydrolysis. There 

K+-ATPase was inactivated by FSB A in the presence of Mg23-. Although Na+ and K+ are not essential for inactivation, they accelerate the inactivation rate. In the absence of Mg2+, the enzyme was not inactivated by FSB A even in the presence of both Na+ and K+. Sequence study on the FSBA-modified dog kidney enzyme revealed that one of the major labeled sites is a lysyl residue equivalent to Lys-725 of the torpedo enzyme. The simultaneous replacement of both Lys-712 and Lys-713 by Met of SR-ATPase (The two lysyl residues are equivalent to Lys-725 and Lys-726 of the torpedo Na+, K+-ATPase) did not change the Ca2+-transporting activity or the formation of phosphoenzyme intermediate,781 indicating that these lysyl residues are not essential for enzyme activity. 

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Ion transporting ATPases that become autophosphory-lated by the terminal phosphate group of ATP during the ion transporting process... 

Although the sequence identity averaged over the whole length of the molecule is generally low among different P-type ion transport ATPases, the conserved sequences around the phosphate acceptor aspartyl group and in the ATP binding domain are well preserved [30,32,46]. Structure predictions based on the hydropathy plots... 

Reliable information about the transmembrane topology of ion transport ATPases can be obtained only by a combination of predictions based on amino acid sequence... 

H -ATPase catalyzes terminal transphosphorylation in ATP synthesis. The presence of phosphorylated intermediate in this reaction was suggested by kinetic analysis [103-105], and isolation of an acylphosphate phosphoenzyme in ion transporting ATPases, including Ca -ATPase [159], Na, K -ATPase [160] and plasma mem-... 

The last mediator of gastric secretion in the parietal cell is an H+,K+-ATPase (proton or acid pump) which is a member of the phosphorylating class of ion transport ATPases. Hydrolysis of ATP results in ion transport. This chemical reaction induces a conformational change in the protein that allows an electroneutral exchange of cytoplasmic H+ for K+. The pump is activated when associated with a potassium chloride pathway in the canalicular membrane which allows potassium chloride efflux into the extracytoplasmic space, and thus results in secretion of hydrochloric acid at the expense of ATP breakdown. The activity of the pump is determined by the access of K+ on this surface on the pump. In the absence of K+, the cycle stops at the level of the phosphoenzyme [137]. 

Rensing C, Ghosh M and Rosen BP (1999) Families of sofi-metal-ion-transporting ATPases. J Bac-teriol 181 5891-5897. 

Scheiner-Bobis, G., Ion-transporting ATPases as ion channels. Naunyn Schmiedebergs Arch Pharmacol, 1998 357(5) 477 82. 

C Rensing, M Ghosh, BP Rosen. Families of soft-metal-ion-transporting ATPases. J Bacteriol 181 5891-5897, 1999. 

Gottardi CJ, Caplan MJ (1993) Molecular requirements for the cell-surface expression of multisubunit ion-transporting ATPases. J Biol Chem 268 14342-14347... 

J. C. Seou (Aarhus) discovery of the first molecular pump, an ion-transporting enzyme Na+-K+ ATPase. 

It is possible that the stationary-state situations leading to an active ion transport occur only in localized regions of the membrane, i.e., at ATPase molecule units with diameters of about 50 A and a length of 80 A. The vectorial ion currents at locations with a mixed potential and special equipotential lines would appear phenomenologically like ionic channels. If the membrane area where the passive diffusion occurs is large, it may determine the rest potential of the whole cell. 

Tlie Na+/K+-ATPase belongs to the P-type ATPases, a family of more than 50 enzymes that also includes the Ca2+-ATPase of the sarcoplasmic reticulum or the gastric H+/K+-ATPase. P-Type ATPases have in common that during ion transport an aspartyl phos-phointermediate is formed by transfer of the y-phosphate group of ATP to the highly conserved sequence DKTGS/T [1]. 

The Na+/K+-ATPase is the only enzyme known to interact with CTS, which reversibly bind to the extracellular side of the Na+/K+-ATPase at the E2-P conformational state [E2-P ouabain] and inhibit ATP hydrolysis and ion transport (Fig. lb, step 4). 

The Ca transport ATPase of sarcoplasmic reticulum is an intrinsic membrane protein of 110 kDa [8-11] that controls the distribution of intracellular Ca by ATP-dependent translocation of Ca " ions from the cytoplasm into the lumen of the sarcoplasmic reticulum [12-16],... 

The determination of the amino acid sequences of the sarcoplasmic reticulum Ca -ATPase [42] and of the closely related Na, K -ATPase [43,44] have opened a new era in the analysis of ion transport mechanisms. Since 1985, several large families of structurally related ion transport enzymes were discovered [3,34,45-50] that are the products of different genes. Within each family several isoenzymes may be produced from a single gene-product by alternative splicing (Table I). 

Our discussion here will concentrate on the various forms of the Ca " transport ATPases that occur in the sarcoplasmic reticulum of muscle cells of diverse fiber types and in the endoplasmic reticulum of nonmuscle cells (SERCA). The structure of these enzymes will be compared with the Ca transport ATPases of surface membranes (PMCA) [3,29-32,34] and with other ATP-dependent ion pumps that transport Na, K, andH [46,50-52]. 

A well-known example of active transport is the sodium-potassium pump that maintains the imbalance of Na and ions across cytoplasmic membranes. Flere, the movement of ions is coupled to the hydrolysis of ATP to ADP and phosphate by the ATPase enzyme, liberating three Na+ out of the cell and pumping in two K [21-23]. Bacteria, mitochondria, and chloroplasts have a similar ion-driven uptake mechanism, but it works in reverse. Instead of ATP hydrolysis driving ion transport, H gradients across the membranes generate the synthesis of ATP from ADP and phosphate [24-27]. 

An essential requirement for diffusion of Na+ ions is the creation of a concentration gradient for sodium between the filtrate and intracellular fluid of the epithelial cells. This is accomplished by the active transport ofNa+ ions through the basolateral membrane of the epithelial cells (see Figure 19.4). Sodium is moved across this basolateral membrane and into the interstitial fluid surrounding the tubule by the Na+, K+-ATPase pump. As a result, the concentration of Na+ ions within the epithelial cells is reduced, facilitating the diffusion of Na+ ions into the cells across the luminal membrane. Potassium ions transported into the epithelial cells as a result of this pump diffuse back into the interstitial fluid (proximal tubule and Loop of Henle) or into the tubular lumen for excretion in the urine (distal tubule and collecting duct). 

Certain enzymes shown to be present in myelin could be involved in ion transport. Carbonic anhydrase has generally been considered a soluble enzyme and a glial marker but myelin accounts for a large part of the membrane-bound form in brain. This enzyme may play a role in removal of carbonic acid from metabolically active axons. The enzymes 5 -nucleotidase and Na+, K+-ATPase have long been considered specific markers for plasma membranes and are found in myelin at low levels. The 5 -nucleotidase activity may be related to a transport mechanism for adenosine, and Na+, K+-ATPase could well be involved in transport of monovalent cations. The presence of these enzymes suggests that myelin may have an active role in ion transport in and out of the axon. In connection with this hypothesis, it is of interest that the PLP gene family may have evolved from a pore-forming polypeptide [9],... 

In biological systems, therefore, the behavior of Li+ is predicted to be similar to that of Na+ and K+ in some cases, and to that of Mg2+ and Ca2+ in others [12]. Indeed, research has demonstrated numerous systems in which one or more of these cations is normally intrinsically involved, including ion transport pathways and enzyme activities, in which Li+ has mimicked the actions of these cations, sometimes producing inhibitory or stimulatory effects. For example, Li+ can replace Na+ in the ATP-dependent system which controls the transport of Na+ through the endoplasmic reticulum Li+ inhibits the activity of some Mg2+-dependent enzymes in vitro, such as pyruvate kinase and inositol monophosphate phosphatase Li+ affects the activity of some Ca2+-dependent enzymes— it increases the levels of activated Ca2+-ATPase in human erythrocyte membranes ex vivo and inhibits tryptophan hydroxylase. 

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