Literature summary for 7.2.4.2 extracted from

Jockel, P.; Schmid, M.; Choinowski, T.; Dimroth, P.
Essential role of tyrosine 229 of the oxaloacetate decarboxylase beta-subunit in the energy coupling mechanism of the Na(+) pump (2000), Biochemistry, 39, 4320-4326.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli DH5alpha/pSK-GAB	Klebsiella pneumoniae

Protein Variants

Protein Variants	Comment	Organism
C291E	beta subunit, unstable in the absence of Na+, dissociates into an alpha-gamma subcomplex and the beta subunit	Klebsiella pneumoniae
D203N	beta subunit, complete loss of activity	Klebsiella pneumoniae
G337A	beta subunit, complete loss of activity	Klebsiella pneumoniae
G377A	beta subunit, complete loss of activity	Klebsiella pneumoniae
G380A	beta subunit, about 30% of activity	Klebsiella pneumoniae
N373L	beta subunit, almost complete loss of activity	Klebsiella pneumoniae
Q192L	beta subunit, lower pH optimum than wild type enzyme	Klebsiella pneumoniae
R389A	beta subunit, about 10% of activity, significantly decreased Na+ binding	Klebsiella pneumoniae
R389D	beta subunit, almost complete loss of activity	Klebsiella pneumoniae
R389K	beta subunit, about 80% of activity, significantly decreased Na+ binding	Klebsiella pneumoniae
R389L	beta subunit, about 10% of activity, significantly decreased Na+ binding	Klebsiella pneumoniae
S382A	beta subunit, complete loss of activity	Klebsiella pneumoniae
S382C	beta subunit, complete loss of activity	Klebsiella pneumoniae
S382D	beta subunit, about 10% of activity	Klebsiella pneumoniae
S382E	beta subunit, complete loss of activity	Klebsiella pneumoniae
S382N	beta subunit, complete loss of activity	Klebsiella pneumoniae
S382Q	beta subunit, complete loss of activity	Klebsiella pneumoniae
S382T	beta subunit, about 10% of activity	Klebsiella pneumoniae
Y209A	beta subunit, lower pH optimum than wild type enzyme	Klebsiella pneumoniae
Y229F	beta subunit, completely inactive	Klebsiella pneumoniae

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	beta and gamma subunits are integral membrane proteins, alpha subunit is attached to the gamma subunit	Klebsiella pneumoniae	16020	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Na+	two ions per enzyme required for maximum activity, cooperative binding, 20-50 mM protects against tryptic digestion	Klebsiella pneumoniae
Zn2+	binds to the gamma subunit	Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
Oxaloacetate	Klebsiella pneumoniae	one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction	Pyruvate + CO2	-	?

Organism

Organism	UniProt	Comment	Textmining
Klebsiella pneumoniae	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant protein from Escherichia coli	Klebsiella pneumoniae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	specific activity for all mutant enzymes	Klebsiella pneumoniae
45	-	wild type enzyme	Klebsiella pneumoniae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Oxaloacetate	-	Klebsiella pneumoniae	Pyruvate + CO2	-	?
Oxaloacetate	one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction	Klebsiella pneumoniae	Pyruvate + CO2	-	?

Subunits

Subunits	Comment	Organism
trimer	alpha, beta, gamma	Klebsiella pneumoniae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.5	-	Q192L and Y209A mutants	Klebsiella pneumoniae
6.5	7.5	wild type enzyme	Klebsiella pneumoniae

Cofactor

Cofactor	Comment	Organism	Structure
biotin	binds to the C-terminal domain of the alpha-subunit	Klebsiella pneumoniae