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Literature summary extracted from

  • Belisario, D.C.; Rocafull, M.A.; del Castillo, J.R.
    Purification and characterization of the ouabain-sensitive H+/K+-ATPase from guinea-pig distal colon (2010), Arch. Biochem. Biophys., 496, 21-32.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
7.2.2.19 ATP ATP concentrations over 5 mM induce H+/K+-ATPase activity inhibition, suggesting the existence of ATP-regulatory sites Cavia porcellus
7.2.2.19 additional information no inhibition by SCH-28080 and bafilomycin-A Cavia porcellus
7.2.2.19 Ouabain
-
Cavia porcellus
7.2.2.19 vanadate
-
Cavia porcellus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7.2.2.19 additional information
-
additional information kinetics at different pH, overview Cavia porcellus
7.2.2.19 0.0245
-
ATP pH 7.4, temperature not specified in the publication, with optimal concentration of ATP, Mg2+ and K+ Cavia porcellus
7.2.2.19 0.0494
-
ATP pH 7.0, temperature not specified in the publication, with optimal concentration of ATP, Mg2+ and K+ Cavia porcellus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
7.2.2.19 additional information the H+/K+-ATPase and Na+/K+-ATPase, EC 3.6.3.9, are not located at the same sub-cellular fraction. The ouabain-insensitive K+-ATPase resembles better the distribution of the Na+/K+-ATPase, suggesting that can be associated with the colonocyte basolateral plasma membrane Cavia porcellus
-
-
7.2.2.19 plasma membrane apical not basolateral Cavia porcellus 5886
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
7.2.2.19 Cs+ activates Cavia porcellus
7.2.2.19 K+ activates, activation by K+ is independent of the accompanying anion, obtaining maximal activity at about 0.5 mM and pH 7.4, Km is 1.9 mM, not effective at pH 6.6 Cavia porcellus
7.2.2.19 Mg2+ dependent on, optimally at 5 mM, Km is 3.4 mM or 0.5 mM with optimal ATP concentration Cavia porcellus
7.2.2.19 additional information H+/K+-ATPase is Na+-independent, Mg2+-dependent, and activated by K+, Rb+, Cs+, and NH4 + but not by Na+, Li+, or N-methyl-glucamine Cavia porcellus
7.2.2.19 NH4+ activates Cavia porcellus
7.2.2.19 Rb+ activates Cavia porcellus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
7.2.2.19 100000
-
1 * 100000, alpha-subunit, + 1 * 500000, beta-subunit, alpha/beta, SDS-PAGE Cavia porcellus
7.2.2.19 500000
-
1 * 100000, alpha-subunit, + 1 * 500000, beta-subunit, alpha/beta, SDS-PAGE Cavia porcellus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7.2.2.19 ATP + H2O + H+/in + K+/out Cavia porcellus
-
ADP + phosphate + H+/out + K+/in
-
?

Organism

EC Number Organism UniProt Comment Textmining
7.2.2.19 Cavia porcellus
-
male guinea pigs
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
7.2.2.19 phosphoprotein the enzyme is phosphorylated from [32P]-gamma-ATP, forming an acyl-phosphate bond, in an Mg2+-dependent, vanadate-sensitive process, and inhibited by hydroxylamine Cavia porcellus

Purification (Commentary)

EC Number Purification (Comment) Organism
7.2.2.19 native ouabain-sensitive H+/K+-ATPase 106.5fold from isolated guinea-pig distal colon apical plasma membranes by solubilization with SDS, and discontinuous sucrose gradient centrifugation, separation from the ouabain-insensitive K+-ATPase Cavia porcellus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
7.2.2.19 colon distal Cavia porcellus
-
7.2.2.19 colonocyte
-
Cavia porcellus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
7.2.2.19 0.07
-
native ouabain-insensitive H+/K+-ATPase from colonic apical plasma membranes, pH not specified in the publication, temperature not specified in the publication Cavia porcellus
7.2.2.19 0.544
-
native ouabain-sensitive H+/K+-ATPase from colonic apical plasma membranes, pH not specified in the publication, temperature not specified in the publication Cavia porcellus
7.2.2.19 2.045
-
purified native ouabain-sensitive H+/K+-ATPase, pH not specified in the publication, temperature not specified in the publication Cavia porcellus

Storage Stability

EC Number Storage Stability Organism
7.2.2.19 -70°C, purified enzyme, loss of over 90% activity without additive, 65% activity remaining with 25% added, over 95% remaining with addtion of 10% DMSO Cavia porcellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7.2.2.19 ATP + H2O + H+/in + K+/out
-
Cavia porcellus ADP + phosphate + H+/out + K+/in
-
?
7.2.2.19 ATP + H2O + H+/in + K+/out a Na+-independent, ouabain-sensitive H+/K+-exchange. K+ can be replaced by H+ in the enzyme reaction cycle, however pH-induced conformational changes cannot be discarded Cavia porcellus ADP + phosphate + H+/out + K+/in
-
?
7.2.2.19 CTP + H2O + H+/in + K+/out a Na+-independent, ouabain-sensitive H+/K+-exchange, low activity with CTP Cavia porcellus CDP + phosphate + H+/out + K+/in
-
?
7.2.2.19 GTP + H2O + H+/in + K+/out a Na+-independent, ouabain-sensitive H+/K+-exchange, low activity with GTP Cavia porcellus GDP + phosphate + H+/out + K+/in
-
?
7.2.2.19 ITP + H2O + H+/in + K+/out a Na+-independent, ouabain-sensitive H+/K+-exchange, low activity with ITP Cavia porcellus IDP + phosphate + H+/out + K+/in
-
?
7.2.2.19 additional information the enzyme is phosphorylated from [32P]-gamma-ATP, forming an acyl-phosphate bond, in an Mg2+-dependent, vanadate-sensitive process Cavia porcellus ?
-
?
7.2.2.19 UTP + H2O + H+/in + K+/out a Na+-independent, ouabain-sensitive H+/K+-exchange, low activity with UTP Cavia porcellus UDP + phosphate + H+/out + K+/in
-
?

Subunits

EC Number Subunits Comment Organism
7.2.2.19 dimer 1 * 100000, alpha-subunit, + 1 * 500000, beta-subunit, alpha/beta, SDS-PAGE Cavia porcellus
7.2.2.19 More H+/K+-ATPase is an alpha/beta-heterodimer, whose subunits, identified by Tandem-mass spectrometry, seem to correspond to HKalpha2 and Na+/K+-ATPase beta1-subunit, respectively Cavia porcellus

Synonyms

EC Number Synonyms Comment Organism
7.2.2.19 H+/K+-ATPase
-
Cavia porcellus
7.2.2.19 ouabain-sensitive H+/K+-ATPase
-
Cavia porcellus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2.2.19 6.6 7.4 the optimal pH for the ouabain-sensitive ATPase activity in the presence of K+ is pH 7.4, while the optimum pH for this activity, in the absence of K+, is pH 6.6 Cavia porcellus

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
7.2.2.19 0.0016
-
pH 7.4, temperature not specified in the publication Cavia porcellus vanadate
7.2.2.19 0.0017
-
pH 7.4, temperature not specified in the publication Cavia porcellus Ouabain

General Information

EC Number General Information Comment Organism
7.2.2.19 physiological function distal colon absorbs K+ through a Na+-independent, ouabain-sensitive H+/K+-exchange, associated to an apical ouabain-sensitive H+/K+-ATPase Cavia porcellus