Characterization of gastric mucosal membranes. IX. Fractionation and purification of K+-ATPase-containing vesicles by zonal centrifugation and free-flow electrophoresis technique

Saccomani, G.; Stewart, H. B.; Shaw, Denise R.; Lewin, M. J. M.; Sachs, George

doi:10.1016/0005-2736(77)90081-5

Cited by 198 publications

(102 citation statements)

References 23 publications

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“…We used medullary bone of regularly egg-laying hens as a source of tissue because of its high number of osteoclasts. H+-transport properties of bone microsomal membrane fraction were compared with gastric microsomes and kidney microsomes that are known to contain H+K+-ATPase and vacuolar AT- Pase, respectively (Saccomani et al, 1977;Gluck et al, 1984). Bone cell microsomes and kidney microsomes showed almost identical inhibition patterns with NEM and orthovanadate, whereas the microsomes derived from the parietal cells of the gastric mucosa showed profound differences.…”

Section: Discussionmentioning

confidence: 99%

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Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

Väänänen

Karhukorpi

Sundquist

et al. 1990

The Journal of Cell Biology

350

115

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Abstract. Microsomal membrane vesicles prepared either from chicken medullary bone or isolated osteoclasts were shown to have ATP-dependent H+-transport activity. This activity was N-ethylmaleimide-sensitive but resistant to oligomycin and orthovanadate, suggesting a vacuolar-type ATPase. Furthermore, immunological cross-reactivity of 60-and 70-kD osteoclast membrane antigens with Neurospora crassa vacuolar ATPase was observed when analyzed by immunoblotting. Same antibodies labeled only osteoclasts in chicken and rat bone in immunohistochemistry. Immunoelectronmicroscopy localized these antigens in apical membranes of rat osteoclasts and kidney intercalated cells of inner stripe of outer medulla. Pretreatmerit of animals with parathyroid hormone enhanced the immunoreaction in the apical membranes of osteoclasts. No immunoreaction was seen in osteoclasts when antibodies against gastric H+,K+-ATPase were used. These results suggest that osteoclast resorbs bone by secreting protons through vacuolar H+-ATPase.STEOCLASTS are multinucleated giant cells that are responsible for bone resorption. Like secreting epithelial cells, they are polarized when active in bone resorption, showing three distinct specialized cell membrane areas (for review see Vaes et al., 1988). In addition to the basolateral membrane, which is rich in Na+,K+-ATPase (Baron et al., 1986), resorbing osteoclast exhibits a clear zone that mediates the attachment of resorbing cells to the bone matrix. The cytoplasm in the vicinity of this clear zone contains specialized cytoskeletal structures (Holtrop et al., 1974;Lakkakorpi et al., 1989). The third specialized membrane area, the ruffled border, faces the actual bone resorption site on the bone surface. The resorption lacuna underneath the ruffled border membrane is acidic. This has been shown by acridine orange accumulation experiments (Anderson et al., 1986; Bar6n et al., 1985) and direct micropuncture measurements (Fallon, 1984). The acidic pH favors dissolution of the bone mineral. In addition, proteinases active at acid pH and capable of collagen degradation are present in osteoclasts (Vaes et al., 1988; Blair et al., 1986). Enzyme histochemistry suggests the presenc$ of ATPase activity in the plasma membrane of the osteoclast- (Akisaka et al., 1986). Baron et al. (1985) found at the ruffled:border of the osteoclast a 100-kD lysosomal membrane polypeptide that showed immunological similarity to gastric H+,K+-ATPase. Now we report that osteoclasts contain an ATP-dependent proton pump that is clearly different from the gastric proton pump and from the mitochondrial proton pump but shows considerable immunological similarities to the vacuolar type H+-ATPase of Neurospora crassa. Materials and Methods Preparation of Bone MicrosomesBone microsomes were prepared from medullary bone of regularly laying hens. Medullary bone from the tibia and femur was dissected out and immediately homogenized in a medium containing 5 mM Tris pH 7.4, 250 mM sucrose, 1 mM K2CO3, 1 mM DTT, and 1 mM EGTA in a glass-Teflun ...

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Section: Discussionmentioning

confidence: 99%

“…Microsomes from chicken kidney medulla were prepared as described earlier for bovine kidney (Gluck et al, 1984). Gastric vesicles from pig stomach were prepared using the method of Saccomani et al (1977). …”

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confidence: 99%

Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

Väänänen

Karhukorpi

Sundquist

et al. 1990

The Journal of Cell Biology

350

115

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“…This phenomenon is attributable to the transformation of the membrane of tubulovesicles to that of intracellular canaliculi, as the continuity between the membranes of intracellular canaliculi and tubulovesicles has been confirmed in rat parietal cells with ultra-high resolution scanning electron microscopic observations (OGATA and YAMASAKI, 1993;OGATA et al, 1995). On the other hand, molecular biochemistry has found that the gastric hydrogen-potassium-stimulated adenosine triphosphatase (H+-K+ ATPase) or proton pump catalyzes the exchange of luminal K+ versus intracellular H+ (GANSER and FORTE, 1973;SACHS et al, 1976;SACCOMANI et al, 1977). Recently, several antibodies for H+-K+ ATPase of parietal cells have been refined.…”

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confidence: 94%

An Immuno- and Enzyme Cytochemical Study of the H+-K+ ATPase in Human Parietal Cells after Administration of Tetragastrin and Omeprazole.

Kobayashi

Araki

Ando

et al. 1997

Archives of Histology and Cytology

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“…The enzyme catalyses the ATPdependent counter transport of protons and potassium ions across the tubulo-vesicular membranes of the gastric parietal cell [ 11. Until recently the H + /K + -ATPase was thought to consist of a single a-subunit of apparent molecular mass 95,000-100,000 [2], which was phosphorylated on an aspartic acid residue during the catalytic cycle [3]. The entire protein sequences of the porcine (1034 amino acids [4]) and rat (1033 amino acids [5]) a-subunits have been deduced from the nucleotide sequences of the corresponding cDNAs.…”

Section: Introductionmentioning

confidence: 99%

The β‐subunit of the gastric H⁺/K⁺‐ATPase can occur without the α‐subunit

Baldwin¹

1990

FEBS Letters

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Characterization of gastric mucosal membranes. IX. Fractionation and purification of K+-ATPase-containing vesicles by zonal centrifugation and free-flow electrophoresis technique

Cited by 198 publications

References 23 publications

Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

An Immuno- and Enzyme Cytochemical Study of the H+-K+ ATPase in Human Parietal Cells after Administration of Tetragastrin and Omeprazole.

The β‐subunit of the gastric H⁺/K⁺‐ATPase can occur without the α‐subunit

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Characterization of gastric mucosal membranes. IX. Fractionation and purification of K+-ATPase-containing vesicles by zonal centrifugation and free-flow electrophoresis technique

Cited by 198 publications

References 23 publications

Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.

An Immuno- and Enzyme Cytochemical Study of the H+-K+ ATPase in Human Parietal Cells after Administration of Tetragastrin and Omeprazole.

The β‐subunit of the gastric H+/K+‐ATPase can occur without the α‐subunit

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The β‐subunit of the gastric H⁺/K⁺‐ATPase can occur without the α‐subunit