Control of Cell Membrane Ecto-ATPase by Oligomerization State:  Intermolecular Cross-Linking Modulates ATPase Activity†

Biochemistry ◽  
1996 ◽  
Vol 35 (25) ◽  
pp. 8289-8298 ◽  
Author(s):  
James G. Stout ◽  
Terence L. Kirley
Keyword(s):  
Cell Membrane ◽  
Atpase Activity ◽  
Cross Linking

scholarly journals The Mg2+-ATPase of rabbit skeletal-muscle transverse tubule is a highly glycosylated multiple-subunit enzyme

1991 ◽  
Vol 278 (2) ◽  
pp. 375-380 ◽  
Author(s):  
T L Kirley
Keyword(s):  
Skeletal Muscle ◽  
Atpase Activity ◽  
Atp Hydrolysis ◽  
Rabbit Skeletal Muscle ◽  
Cross Linking ◽  
Low Concentrations ◽  
Sds Page ◽  
Molecular Masses ◽  
Relationship Of ◽  
Peptide Core

The Mg(2+)-ATPase present in rabbit skeletal-muscle transverse tubules is an integral membrane enzyme which has been solubilized and purified previously in this laboratory [Kirley (1988) J. Biol. Chem. 263, 12682-12689]. The present study indicates that, in addition to the approx. 100 kDa protein (distinct from the sarcoplasmic-reticulum Ca(2+)-ATPase) seen previously to co-purify with the Mg(2+)-ATPase activity, there are also proteins having molecular masses of 160, 70 and 43 kDa. The 70 and 43 kDa glycosylated proteins (50 and 31 kDa after deglycosylation) are difficult to detect by SDS/PAGE before deglycosylation, owing to the broadness of the bands. Additional purification procedures, cross-linking studies and chemical and enzymic deglycosylation studies were undertaken to determine the structure and relationship of these proteins. Both the 97 and 160 kDa proteins were demonstrated to be N-glycosylated at multiple sites, the 97 kDa protein being reduced to a peptide core of 84 kDa and the 160 kDa protein to a peptide core of 131 kDa after deglycosylation. Although the Mg(2+)-ATPase activity is resistant to a number of chemical modification reagents, cross-linking inactivates the enzyme at low concentrations. This inactivation is accompanied by cross-linking of two 97 kDa molecules to one another, suggesting that the 97 kDa protein is involved in ATP hydrolysis. The existence of several proteins along with the inhibition of ATPase activity by cross-linking is consistent with the interpretation of the susceptibility of this enzyme to inactivation by most detergents as being due to the disruption of a protein complex of associated subunits by the inactivating detergents. The 160 kDa glycoprotein can be partially resolved from the Mg(2+)-ATPase activity, and is identified by its N-terminal amino acid sequence as angiotensin-converting enzyme.

scholarly journals Dissociation of clathrin coats coupled to the hydrolysis of ATP: role of an uncoating ATPase.

1984 ◽  
Vol 99 (2) ◽  
pp. 734-741 ◽  
Author(s):  
W A Braell ◽  
D M Schlossman ◽  
S L Schmid ◽  
J E Rothman
Keyword(s):  
Atpase Activity ◽  
Atp Hydrolysis ◽  
Low Ph ◽  
Coated Vesicles ◽  
Magnesium Concentration ◽  
Cross Linking ◽  
Dependent Atpase ◽  
Event Triggering ◽  
Hydrolysis Of

ATP hydrolysis was used to power the enzymatic release of clathrin from coated vesicles. The 70,000-mol-wt protein, purified on the basis of its ATP-dependent ability to disassemble clathrin cages, was found to possess a clathrin-dependent ATPase activity. Hydrolysis was specific for ATP; neither dATP nor other ribonucleotide triphosphates would either substitute for ATP or inhibit the hydrolysis of ATP in the presence of clathrin cages. The ATPase activity is elicited by clathrin in the form of assembled cages, but not by clathrin trimers, the product of cage disassembly. The 70,000-mol-wt polypeptide, but not clathrin, was labeled by ATP in photochemical cross-linking, indicating that the hydrolytic site for ATP resides on the uncoating protein. Conditions of low pH or high magnesium concentration uncouple ATP hydrolysis from clathrin release, as ATP is hydrolyzed although essentially no clathrin is released. This suggests that the recognition event triggering clathrin-dependent ATP hydrolysis occurs in the absence of clathrin release, and presumably precedes such release.

scholarly journals Intracellular Na+ Controls Cell Surface Expression of Na,K-ATPase via a cAMP-independent PKA Pathway in Mammalian Kidney Collecting Duct Cells

2003 ◽  
Vol 14 (7) ◽  
pp. 2677-2688 ◽  
Author(s):  
Manlio Vinciguerra ◽  
Georges Deschênes ◽  
Udo Hasler ◽  
David Mordasini ◽  
Martine Rousselot ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Cell Surface ◽  
Atpase Activity ◽  
Collecting Duct ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Cortical Collecting Duct ◽  
Mammalian Kidney ◽  
Duct Cells ◽  
Collecting Duct Cells

In the mammalian kidney the fine control of Na+ reabsorption takes place in collecting duct principal cells where basolateral Na,K-ATPase provides the driving force for vectorial Na+ transport. In the cortical collecting duct (CCD), a rise in intracellular Na+ concentration ([Na+]i) was shown to increase Na,K-ATPase activity and the number of ouabain binding sites, but the mechanism responsible for this event has not yet been elucidated. A rise in [Na+]i caused by incubation with the Na+ ionophore nystatin, increased Na,K-ATPase activity and cell surface expression to the same extent in isolated rat CCD. In cultured mouse mpkCCDcl4 collecting duct cells, increasing [Na+]i either by cell membrane permeabilization with amphotericin B or nystatin, or by incubating cells in a K+-free medium, also increased Na,K-ATPase cell surface expression. The [Na+]i-dependent increase in Na,K-ATPase cell-surface expression was prevented by PKA inhibitors H89 and PKI. Moreover, the effects of [Na+]i and cAMP were not additive. However, [Na+]i-dependent activation of PKA was not associated with an increase in cellular cAMP but was prevented by inhibiting the proteasome. These findings suggest that Na,K-ATPase may be recruited to the cell membrane following an increase in [Na+]i through cAMP-independent PKA activation that is itself dependent on proteasomal activity.

Stimulatory effect of vanadate on (Na+ + K+)-ATPase activity and on 3H-ouabain-binding in a cat heart cell membrane preparation

Nature ◽  
1979 ◽  
Vol 278 (5703) ◽  
pp. 459-461 ◽  
Author(s):  
ERLAND ERDMANN ◽  
WOLFGANG KRAWIETZ ◽  
GUNTHER PHILIPP ◽  
INGELORE HACKBARTH ◽  
WILHELM SCHMITZ ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Atpase Activity ◽  
Membrane Preparation ◽  
Heart Cell ◽  
Ouabain Binding ◽  
Heart Cell Membrane ◽  
Cell Membrane Preparation ◽  
Cat Heart

Cell membrane calcium ATPase activity in hypertension

1987 ◽  
Vol 15 (5) ◽  
pp. 854-855
Author(s):  
ABIMBOLA ADEOYA ◽  
ROBERT I. NORMAN ◽  
ROBERT F. BING
Keyword(s):  
Cell Membrane ◽  
Atpase Activity ◽  
Calcium Atpase

scholarly journals LIGAND-INDUCED MOVEMENT OF LYMPHOCYTE MEMBRANE MACROMOLECULES

1973 ◽  
Vol 137 (3) ◽  
pp. 675-689 ◽  
Author(s):  
Emil R. Unanue ◽  
Morris J. Karnovsky ◽  
Howard D. Engers
Keyword(s):  
Protein Synthesis ◽  
Cell Membrane ◽  
Cell Surface ◽  
Oxidative Phosphorylation ◽  
Cross Linking ◽  
Lymphocyte Membrane ◽  
Compact Zone ◽  
Entire Cell ◽  
Freeze Etching ◽  
Spleen Lymphocytes

Spleen lymphocytes were studied for the movement and interiorization of complexes of anti-Ig-surface Ig. The movement of the complex into a small, compact zone of the cell membrane (forming a cap) was inhibited by drugs that inhibited glycolysis and oxidative phosphorylation, but not by drugs that affected protein synthesis. Dead lymphocytes did not form caps. Freeze-etching techniques revealed that inhibited lymphocytes showed formation of multiple small complexes over the entire cell surface. Inhibitors of glycolysis and of oxidative phosphorylation also inhibited the interiorization and catabolism of radioiodinated anti-Ig. We hypothesize that cross-linking of all the surface Ig triggers the membrane movements that are required to pull the lattice into one zone of the cell.

Flotillin-mediated endocytosis and ALIX–syntenin-1–mediated exocytosis protect the cell membrane from damage caused by necroptosis

2019 ◽  
Vol 12 (583) ◽  
pp. eaaw3423 ◽  
Author(s):  
Weiliang Fan ◽  
Jia Guo ◽  
Beichen Gao ◽  
Wenbin Zhang ◽  
Liucong Ling ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Membrane ◽  
Lipid Raft ◽  
Affinity Purification ◽  
Human Diseases ◽  
Membrane Disruption ◽  
Cross Linking ◽  
Lysosomal Degradation ◽  
Regulated Necrosis ◽  
Associated Proteins

Necroptosis is a form of regulated necrosis that is implicated in various human diseases including Alzheimer’s disease. Necroptosis requires the translocation of the pseudokinase MLKL from the cytosol to the plasma membrane after its phosphorylation by the kinase RIPK3. Using protein cross-linking followed by affinity purification, we detected the lipid raft–associated proteins flotillin-1 and flotillin-2 and the ESCRT-associated proteins ALIX and syntenin-1 in membrane-localized MLKL immunoprecipitates. Phosphorylated MLKL was removed from membranes through either flotillin-mediated endocytosis followed by lysosomal degradation or ALIX–syntenin-1–mediated exocytosis. Thus, cells undergoing necroptosis need to overcome these independent suppressive mechanisms before plasma membrane disruption can occur.

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