Information on EC 7.2.2.12 - P-type Zn2+ transporter

-

Substrates: -
Products: -

?

ATP + H2O + Ca2+/out

ADP + phosphate + Ca2+/in

-

Substrates: -
Products: -

?

ATP + H2O + Cd(thiolate)2 [side 1]

ADP + phosphate + Cd(thiolate)2 [side 2]

-

Substrates: -
Products: -

?

ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/out

ATP + H2O + Cd2+/out

ADP + phosphate + Cd2+/in

ATP + H2O + Cd2+[side 1]

ADP + phosphate + Cd2+[side 2]

ATP + H2O + Co2+/in

ADP + phosphate + Co2+/out

ATP + H2O + Cu2+/in

ADP + phosphate + Cu2+/out

ATP + H2O + Cu2+/out

ADP + phosphate + Cu2+/in

ATP + H2O + Fe3+/out

ADP + phosphate + Fe3+/in

-

Substrates: -
Products: -

?

ATP + H2O + Hg2+/in

ADP + phosphate + Hg2+/out

-

Substrates: -
Products: -

?

ATP + H2O + Mn2+/out

ADP + phosphate + Mn2+/in

-

Substrates: -
Products: -

?

ATP + H2O + Ni2+/in

ADP + phosphate + Ni2+/out

ATP + H2O + Pb(thiolate)2 [side 1]

ADP + phosphate + Pb(thiolate)2 [side 2]

-

Substrates: -
Products: -

?

ATP + H2O + Pb2+/in

ADP + phosphate + Pb2+/out

9 entries

ATP + H2O + Pb2+[side 1]

ADP + phosphate + Pb2+[side 2]

ATP + H2O + Zn(thiolate)2 [side 1]

ADP + phosphate + Zn(thiolate)2 [side 2]

-

Substrates: -
Products: -

?

ATP + H2O + Zn2+/in

ADP + phosphate + Zn2+/out

25 entries

ATP + H2O + Zn2+/out

ADP + phosphate + Zn2+/in

ATP + H2O + Zn2+[side 1]

ADP + phosphate + Zn2+[side 2]

additional information

?

-

ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/out

ATP + H2O + Cd2+/out

ADP + phosphate + Cd2+/in

-

684877

Substrates: -
Products: -

?

ATP + H2O + Cd2+[side 1]

ADP + phosphate + Cd2+[side 2]

ATP + H2O + Co2+/in

ADP + phosphate + Co2+/out

-

Substrates: -
Products: -

?

ATP + H2O + Cu2+/in

ADP + phosphate + Cu2+/out

-

Substrates: -
Products: -

?

ATP + H2O + Cu2+/out

ADP + phosphate + Cu2+/in

-

Substrates: -
Products: -

?

ATP + H2O + Fe3+/out

ADP + phosphate + Fe3+/in

-

Substrates: -
Products: -

?

ATP + H2O + Mn2+/out

ADP + phosphate + Mn2+/in

-

Substrates: -
Products: -

?

ATP + H2O + Ni2+/in

ADP + phosphate + Ni2+/out

-

Substrates: -
Products: -

?

ATP + H2O + Pb2+/in

ADP + phosphate + Pb2+/out

4 entries

ATP + H2O + Zn2+/in

ADP + phosphate + Zn2+/out

ATP + H2O + Zn2+/out

ADP + phosphate + Zn2+/in

ATP + H2O + Zn2+[side 1]

ADP + phosphate + Zn2+[side 2]

additional information

?

-

Pseudomonas putida

-

657408

Substrates: The enzyme mediates active extrusion of Zn2+, which occurred during the exponential phase of growth. The effluxed Zn2+ are not ejected out of the cell but stored in the outer membrane and periplasm.
Products: -

?

ATP

-

700485, 700757

Ag+

-

activates ATPase activity

Cd2+

Cs2+

-

ZntA confers resistance specifically to Pb2+, Zn2+, and Cd2+ in Escherichia coli. Wild-type ZntA binds two metal ions with high affinity, one in the N-terminal domain and another in the transmembrane domain

Cu2+

Pb2+

Zn2+

additional information

Cd2+

Cu2+

nanobody Nb9

more than 50% inhibition of the ATPase activity

765999

-

thapsigargin

-

vanadate

additional information

no inhibition at 10 mM NPE-caged ATP

750071

-

1,2-dimyristoyl-phosphatidylglycerol

-

solubilized ZntA is increased in the presence of negatively charged phospholipids and at slightly acidic pH. Among the most abundant naturally accuring phospholipids, only phosphatidyl-glycerol enhances the in vitro ATPase activity of TntA. Relipidation of detergent-purified ZntA with 1,2-dioleoylphosphatidyl-glycerol increases the ATPase activity 4fold compared to the purified state. Among the phosphatidylglycerol family, highest activity is observed for 1,2-dioleoyl-phosphatidylglycerol followed by 1,2-dimyristoyl-phosphatidylglycerol, 1,2-dipalmitoyl-phosphatidylglycerol and 1,2-distearoyl-phosphatidylglycerol

1,2-dioleoyl-phosphatidylglycerol

-

solubilized ZntA is increased in the presence of negatively charged phospholipids and at slightly acidic pH. Among the most abundant naturally accuring phospholipids, only phosphatidyl-glycerol enhances the in vitro ATPase activity of TntA. Relipidation of detergent-purified ZntA with 1,2-dioleoylphosphatidyl-glycerol increases the ATPase activity 4fold compared to the purified state. Among the phosphatidylglycerol family, highest activity is observed for 1,2-dioleoyl-phosphatidylglycerol followed by 1,2-dimyristoyl-phosphatidylglycerol, 1,2-dipalmitoyl-phosphatidylglycerol and 1,2-distearoyl-phosphatidylglycerol

1,2-dipalmitoyl-phosphatidylglycerol

-

solubilized ZntA is increased in the presence of negatively charged phospholipids and at slightly acidic pH. Among the most abundant naturally accuring phospholipids, only phosphatidyl-glycerol enhances the in vitro ATPase activity of TntA. Relipidation of detergent-purified ZntA with 1,2-dioleoylphosphatidyl-glycerol increases the ATPase activity 4fold compared to the purified state. Among the phosphatidylglycerol family, highest activity is observed for 1,2-dioleoyl-phosphatidylglycerol followed by 1,2-dimyristoyl-phosphatidylglycerol, 1,2-dipalmitoyl-phosphatidylglycerol and 1,2-distearoyl-phosphatidylglycerol

1,2-distearoyl-phosphatidylglycerol

-

solubilized ZntA is increased in the presence of negatively charged phospholipids and at slightly acidic pH. Among the most abundant naturally accuring phospholipids, only phosphatidyl-glycerol enhances the in vitro ATPase activity of TntA. Relipidation of detergent-purified ZntA with 1,2-dioleoylphosphatidyl-glycerol increases the ATPase activity 4fold compared to the purified state. Among the phosphatidylglycerol family, highest activity is observed for 1,2-dioleoyl-phosphatidylglycerol followed by 1,2-dimyristoyl-phosphatidylglycerol, 1,2-dipalmitoyl-phosphatidylglycerol and 1,2-distearoyl-phosphatidylglycerol

Cd2+

L-cysteine

-

activates the enzyme and mutant lacking 100 residues

Phospholipid

-

solubilized ZntA is increased in the presence of negatively charged phospholipids and at slightly acidic pH. Among the most abundant naturally accuring phospholipids, only phosphatidyl-glycerol enhances the in vitro ATPase activity of TntA. Relipidation of detergent-purified ZntA with 1,2-dioleoylphosphatidyl-glycerol increases the ATPase activity 4fold compared to the purified state. Among the phosphatidylglycerol family, highest activity is observed for 1,2-dioleoyl-phosphatidylglycerol followed by 1,2-dimyristoyl-phosphatidylglycerol, 1,2-dipalmitoyl-phosphatidylglycerol and 1,2-distearoyl-phosphatidylglycerol

Zn2+

additional information

-

stimulated by an outwardly directed proton gradient

-

0.0026 - 0.075

ATP

0.115

Cd(thiolate)2

-

0.0031 - 0.152

Cd2+

18 entries

0.004 - 0.133

Co2+

14 entries

0.0034 - 0.171

Cu2+

14 entries

0.0021 - 0.169

Ni2+

0.118

Pb(thiolate)2

-

0.005 - 0.15

Pb2+

0.0048

Pb2+[side 1]

P37617

pH 7.0, 37°C, wild-type enzyme

766207

-

0.109

Zn(thiolate)2

-

0.0051 - 0.2288

Zn2+

26 entries

0.0103

Zn2+[side 1]

P37617

pH 7.0, 37°C, wild-type enzyme

766207

300

ATP

Q70LF4

in the presence of Zn2+ or Cd2+, in 0.6 M Tris, 2 M NaCl and 0.1 M MgCl2, pH 8.0, at 30°C

718974

0.0681

Cd2+

-

0.0327

Cu2+

-

0.001242

-

0.001584

-

pH condition equal, pH = 7.0/7.0

0.001668

-

pH condition equal, pH = 5.0/5.0

0.001866

-

pH condition outward gradient, pH = 5.0/7.0

0.001916

-

carrier transport is stimulated by the addition of 1 mM ATP

0.002082

-

pH condition inward gradient, pH = 7.0/5.0

additional information

-

The activity with Ni2+, Co2+ and Cu2+ are extremely low, 10-20fold lower compared with the values obtained with Pb2+, Zn2+ and Cd2+.

656669

5.8

-

Cd(II)-dependent activity

6

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ATPase activity assay

6 - 6.5

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Pb(II)-dependent and Zn(II)-dependent activity

6.7

-

7

7.2

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Zn2+ transport measurement at room temperature

7.5

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ATPase activity assay

685173, 687525

23

-

Zn2+ transport measurement at room temperature

30

37

42

-

additional information

-

phosphorylation assays are carried out on ice and room temperature

Agrobacterium tumefaciens

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734648

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Agrobacterium tumefaciens C58 / ATCC 33970

-

-

Cupriavidus metallidurans

Cupriavidus metallidurans ATCC 43123

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767058

Q1LEH0

UniProt

-

751181

-

Geobacillus stearothermophilus

Escherichia coli BW25113

-

-

cf. EC 7.2.2.21

SwissProt

Salmonella enterica serovar typhimurium cells expressing the cadA gene of Geobacillus stearothermophilus

685477

-

-

Salmonella enterica subsp. enterica serovar Typhimurium

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Klebsiella pneumoniae

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767442

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Klebsiella pneumoniae AJ218

-

-

-

-

-

UniProt

Oryza sativa MTU 7029

-

-

-

-

-

-

-

-

-

-

-

UniProt

Shigella sonnei Ss046

-

750071, 765999

UniProt

-

-

Thermus thermophilus

-

719741

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Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039

-

-

-

-

-

-

-

-

-

-

-

-

-

-

additional information

basolateral plasma membrane

-

expression pattern of HMA1 invarious tissues of wild-type plants, HMA1 is expressed preferentially in shoots, including rosette leaves, cauline leaves, ?owers and stems, but little expression is observed in the roots, overview

700757

-

-

lipoprotein, detected by antibodies

-

-

isoform ZntB is the main Zn2+ transporter of the endosome

5768

751181

Golgi apparatus

-

-

lysosomal membrane vesicle, LMV

5765

lysosome

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isoform ZntB is the main Zn2+ transporter of the lysosome

-

isoform ZntA is the main Zn2+ transporter of the contractile vacuole

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Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

malfunction

physiological function

21 entries

128000

-

predicted protein size

684877

160000

-

dimeric form, SDS-PAGE

656669

36000

-

lipoprotein, SDS-PAGE

50000

-

second subunit that contains the phosphorylation site, determined by SDS-PAGE and Western-blotting

60000

60000 - 70000

69000

-

the mutant lacking the N-terminal domain and both the CCCDXXC and GXXCXXC, SDS-PAGE

80000

4 entries

92000

additional information

-

on PAA gels the protein is present in three forms, as a monomer with an apparent mass of 92 kDa, as a dimer of 190 kDa and as a proteolytic fragment of 67 kDa

?

x * 60000, SDS-PAGE

dimer

monomer

additional information

5 entries

C-MBD

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HMA2 C-terminal metal binding domain

C17A

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mutant, 36% decrease in Vmax

C18A

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mutant, 34% decrease in Vmax

C357G

-

mutant enzyme does not confer Cd and Zn resistance to yeast

668627

D149A

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the mutant shows about 135% of wild type Zn2+-transporting activity

D181A

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the mutant shows wild type Zn2+-transporting activity

D313A

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the mutant shows about 130% of wild type Zn2+-transporting activity

D688A

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the mutant shows about 20% of wild type Zn2+-transporting activity

DELTAC-HMA2

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mutant, lacking the 244 C-terminal amino acids

DELTAN-HMA2

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mutant, lacking the N-terminal first 75 amino acids, 56% decrease in Vmax

DELTANC-HMA2

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mutant, lacking the N-terminal first 75 amino acids and the 244 C-terminal amino acids

E169A

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the mutant shows about 25% of wild type Zn2+-transporting activity

E21A

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mutant, 43% decrease in Vmax

E21C

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mutant, 60% decrease in Vmax

F177A

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the mutant shows about 95% of wild type Zn2+-transporting activity

F177L

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the mutant shows about 40% of wild type Zn2+-transporting activity and reduced Cd2+-transporting activity (about 20%) compared to the wild type enzyme

G360A

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the mutant shows about 75% of wild type Zn2+-transporting activity

K138A

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the mutant shows about 110% of wild type Zn2+-transporting activity

K667A

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the mutant shows about 30% of wild type Zn2+-transporting activity

N151A

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the mutant shows about 70% of wild type Zn2+-transporting activity

P366L

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the mutant shows about 15% of wild type Zn2+-transporting activity

S20A

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mutant, shows Vmax similar to that of wild-type HMA2

S20C

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mutant, shows Vmax similar to that of wild-type HMA2

V154A

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the mutant shows about 60% of wild type Zn2+-transporting activity

V154S

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the mutant shows about 60% of wild type Zn2+-transporting activity

A508F

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at low ATP concentrations the mutant enzyme is poorly phosphorylated

C392A

C392A/C394A

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mutant enzyme binds metal ions only at the N-terminal site

C392H

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mutant for evaluating the importance of the cysteine residue of the conserved 392CPC394 motif in metal ion binding

C392H/C394H

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mutant for evaluating the importance of the cysteine residue of the conserved 392CPC394 motif in metal ion binding

C392S

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mutant for evaluating the importance of the cysteine residue of the conserved 392CPC394 motif in metal ion binding

C392S/C394S

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mutant for evaluating the importance of the cysteine residue of the conserved 392CPC394 motif in metal ion binding

C394A

C394S

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mutant for evaluating the importance of the cysteine residue of the conserved 392CPC394 motif in metal ion binding

C59A/C62A

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mutant enzyme in which the N-terminal metal-binding site is disabled by site-specific mutagenesis, can bind only one metal ion

D436N

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the mutant is completely inactive with respect to both in vivo resistance and ATP hydrolysis activity

656669

D714A

D714E

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mutant enzyme is not able to confer resistance to Pb2+, Zn2+, and Cd2+ salts, large reduction in ATPase activity, retains the ability to bind metal ions with high affinity at the transmembrane site

667728

D714H

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mutant enzyme is not able to confer resistance to Pb2+, Zn2+, and Cd2+ salts, large reduction in ATPase activity, retains the ability to bind metal ions with high affinity at the transmembrane site

667728

D714M

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significant metal-independent ATPase activity, poor phosphorylation by ATP and Pi

D714P

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mutant enzyme is not able to confer resistance to Pb2+, Zn2+, and Cd2+ salts, large reduction in ATPase activity

667728

E470A

-

metal ion-stimulated activity is reduced, about 30-40%. Mutant is phosphorylated more strongly in comparison to the wild type enzyme

G444V

-

mutation stabilizes the E2-P state. Gly144 might become close to P upon domain movements

G503S

-

at low ATP concentrations the mutant enzyme is poorly phosphorylated. The phosphorylation defect of the mutant enzyme can be partially compensated by using higher ATP concentrations

G505R

-

at low ATP concentrations the mutant enzymeis poorly phosphorylated. The phosphorylation defect of the mutant enzyme can be fully compensated by using higher ATP concentrations

H475A

-

mutant enzyme reacts poorly with ATP, mutation influences the binding of ATP and also other catalytic steps

H475D

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mutant enzyme reacts poorly with ATP, mutation influences the binding of ATP and also other catalytic steps

H475L

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mutant enzyme reacts poorly with ATP, mutation influences the binding of ATP and also other catalytic steps

H475Q

-

metal ion-stimulated activity is reduced, about 30-40%. Mutant is phosphorylated more weakly in comparison to the wild type enzyme. Mutation affects the reaction with ATP and phosphate and stabilizes the enzyme in a dephosphorylated state

H475S

-

mutant enzyme reacts poorly with ATP, mutation influences the binding of ATP and also other catalytic steps

K693N

-

very low activity, no Cu2+-dependent phosphorylation with ATP, hyperphosphorylation by Pi, altered metal sensitivity of phosphorylation by Pi

K693N/D714M

-

very low activity and phosphorylation with ATP, almost normal phosphorylation with Pi, altered metal senstivity of Pi phosphorylation

P393A

-

mutant, shows no activity and can not bind any metal ion at the transmembrane site

SAAS

-

mutation of two the two cysteines to serines in the CAAC motif near the N-terminus, ATPase activity 50%, diminished phosphorylation, faster dephosphorylation

SPS

-

mutation of two the two cysteines to serines in the CPC motif in transmembrane helix 6, inactive, very low phosphorylation

C392A

Escherichia coli K12

-

mutant DELTA231-ZntA(C392A), binds Pb2+ and Zn2+ with a stoichiometry of 0.5 and with similar or slightly higher affinity than DELTA231-ZntA. Cd2+ does not bind to the mutant enzyme

-

C394A

Escherichia coli K12

-

mutant DELTA231-ZntA(C394A), binds Pb2+ and Zn2+ with a stoichiometry of 0.5 and with similar or slightly higher affinity than DELTA231-ZntA. The mutant enzyme binds Cd2+ with unchanged affinity and with a stoichiometry of 0.5

-

D714A

Escherichia coli K12

-

mutant DELTA231-ZntA(D714A), binds Pb2+ and Zn2+ with a stoichiometry of 0.5 and with similar or slightly higher affinity than DELTA231-ZntA. Cd2+ does not bind to the mutant enzyme

-

additional information

9 entries

the enzyme requires the presence of 20% (v/v) glycerol as stabilizing agent. The enzyme activity requires addition of lipids like soybean extract.

750071

-70°C, 50 mM Tris-HCl, pH 7.5, 300 mM NaCl, 20% glycerol, 2 mM beta-mercaptoethanol, 0.5 mM PMSF

-

-80°C, 100 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM Tris(2-carboxyethyl)phosphine hydrochoride

-

-

isolated from roots, purified via metal affinity chromatography

-

684877

lipoprotein MtsA

-

lysosomal membrane vesicles are prepared

-

membranes from yeast are prepared, purification of the HMA2 C-terminal metal binding domain is performed using a Strep-Tactin Superflow column

-

mutant enzymes C59A/C62A and C392A/C394A

-

Ni-IDA metal affinity column chromatography, gel filtration

Q70LF4

718974

Ni2+-nitrilotriacetic acid resin column chromatography

-

718990

purification of the HMA2 N-terminal metal binding domain is performed using Strep-tag affinity chromatography, membranes from HMA2 expressing yeasts are prepared

-

Talon resin column chromatography

Thermus thermophilus

-

719741

the membrane fractions of the cells are isolated

-

the recombinant mutant lacking the N-terminal domain and both the CCCDXXC and GXXCXXC

-

using a Talon-affinity column

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Zn2+ affinity column chromatography

-

-

Arabidopsis thaliana HMA2 and truncated forms of the protein are prepared using the bacterial expression vector pPRIBA1 and the yeast expression vector pYES2/CT

-

expressed in Arabidopsis thaliana roots and shoots

Triticum aestivum

-

751854

expressed in Escherichia coli BL21(DE3) cells

750071

expressed in Escherichia coli C43 (DE3) cells

Thermus thermophilus

-

719741

expressed in Escherichia coli Top10 cells

-

718990

expression in Escherichia coli

Proteus mirabilis

-

246923

expression in Escherichia coli C43

765999

expression in Saccharomyces cerevisiae

Q9SZW4

670581

expression of wild-type HMA2 and mutant derivatives in transgenic hma2,hma4, Zn-deficient plants from the HMA2 promoter, quantitative realtime PCR expression analysis

-

700485

HMA1 DNA and amino acid sequence determination and analysis, phylogenetic analysis, quantitative realtime RT-PCR expression analysis, expression in Saccharomyces cerevisiae

-

700757

identification of two independent mutant alleles for both genes, HMA2 and HMA4. The hma2-2 mutant is indistinguishable from the wild-type, whereas the hma4-1 and the double mutant hma2-2,hma4-1 accumulate approximately 2fold and 4fold less Zn, respectively, than the wild type.

-

656971

into the pBAD/Myc-Hic C or pBAD/Myc-His A vector for expression in the Escherichia coli strain LMG194(zntA::cat)

-

into the pTRCHisA vector for expression in the Escherichia coli TOP 10 strain

-

mtsA gene

-

mutant enzymes C59A/C62A and C392A/C394A

-

Cupriavidus metallidurans

The cadA and zntA genes are located on the bacterial chromosome. Expression of zntA in Escherichia coli predominantly mediated resistance to zinc, and expression of cadA predominantly mediated resistance to cadmium.

-

655856

the PCR-amplified cadA gene from Geobacillus stearothermophilus is cloned into the expression vector pBAD-TOPO

Geobacillus stearothermophilus

-

685477

the yeast expression vector pYES2/CT carrying Arabidopsis thaliana HMA2 containing a C-terminal Strep-tag is prepared, the HMA2 N-terminal metal binding domain is cloned into the pPRIBA1 vector for expression in Escherichia coli BL21DE3pLysS cells

-

after zinc shock (0.05 mM Zn2+), the mRNA level increases substantially in a zinc-dependent manner

expression of the zntA gene is inducible by CdCl2, ZnCl2 and CoCl2, of which CdCl2 is the most potent inducer. The metal-induced expression of zntA is controlled by the MerR-like regulator ZntR

in roots, isoform HMA4 is up-regulated in response to deficient Zn2+. In roots the isoform HMA4 expression is enhanced only in excess of metal (either Cd2+ orZn2+)

organic acid supply enhances carboxylic speciation of Cd which in turn favors vacuolar sequestration of Cd in the root along with the overexpression of OsHMA3

there is no significant difference in HMA2 expression after Cd2+ or Zn2+ treatment

Oryza sativa

E7EC32

720659

treatment with 18 microM of zinc for 1 h, increases NpunR4017 mRNA levels by up to 1300% above basal levels

Nostoc punctiforme

B2J6J1

733192

upregulated in response to cellular zinc stress