Information on EC 7.2.2.2 - ABC-type Cd2+ transporter

ATP phosphohydrolase (ABC-type, heavy-metal-exporting)

An ATP-binding cassette (ABC) type transporter, characterized by the presence of two similar ATP-binding domains/proteins and two integral membrane domains/proteins. Does not undergo phosphorylation during the transport process. A yeast enzyme that exports some heavy metals, especially Cd2+, from the cytosol into the vacuole.

9000-83-3

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ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/in

-

Substrates: ABC transporter, HMT-1, confers tolerance to cadmium. HMT-1 is not essential for vacuolar phytochelatin sequestration. HMT-1 either does not transport Cd-PC complexes or is not the principal Cd-PC/apoPC transporter
Products: -

?

ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/out

15 entries

ATP + H2O + Cd2+[side 1]

ADP + phosphate + Cd2+[side 2]

ATP + H2O + Co2+[side 1]

ADP + phosphate + Co2+[side 2]

Substrates: -
Products: -

?

ATP + H2O + estradiol-beta-17-glucuronide [side 1]

ADP + phosphate + estradiol-beta-17-glucuronide [side 2]

-

Substrates: -
Products: -

?

ATP + H2O + leukotriene C4 [side 1]

ADP + phosphate + leukotriene C4 [side 2]

-

Substrates: -
Products: -

?

ATP + H2O + Zn2+[side 1]

ADP + phosphate + Zn2+[side 2]

Substrates: lowest activity
Products: -

?

bilirubin/cytoplasm + ATP + H2O

bilirubin/vacuole + ADP + phosphate

-

Substrates: unconjugated substrate
Products: -

?

Cd x glutathione S2/cytoplasm + ATP + H2O

Cd x glutathione S2/vacuole + phosphate

-

Substrates: -
Products: -

?

Cd/cytoplasm + ATP + H2O

Cd/vacuole + ADP + phosphate

Cd2+/cytoplasm + ATP + H2O

Cd2+/vacuole + ADP + phosphate

-

Substrates: -
Products: -

?

diazaborine/cytoplasm + ATP + H2O

diazaborine/vacuole + ADP + phosphate

-

289189

Substrates: -
Products: -

?

glutathione S-conjugated leukotriene C4/cytoplasm + ATP + H2O

glutathione S-conjugated leukotriene C4/vacuole + ADP + phosphate

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289187, 289190, 289186

Substrates: -
Products: -

?

glutathione S-conjugates/cytoplasm + ATP + H2O

glutathione S-conjugates/vacuole + ADP + phosphate

-

Substrates: -
Products: -

?

glutathione/cytoplasm + ATP + H2O

glutathione/vacuole + ADP + phosphate

-

Substrates: -
Products: -

?

S-(2,4-dinitrophenyl)glutathione/cytoplasm + ATP + H2O

S-(2,4-dinitrophenyl)glutathione/vacuole + ADP + phosphate

-

289186, 289187, 289191

Substrates: -
Products: -

?

additional information

?

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ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/in

-

Substrates: ABC transporter, HMT-1, confers tolerance to cadmium. HMT-1 is not essential for vacuolar phytochelatin sequestration. HMT-1 either does not transport Cd-PC complexes or is not the principal Cd-PC/apoPC transporter
Products: -

?

ATP + H2O + Cd2+/in

ADP + phosphate + Cd2+/out

7 entries

ATP + H2O + Cd2+[side 1]

ADP + phosphate + Cd2+[side 2]

ATP + H2O + Co2+[side 1]

ADP + phosphate + Co2+[side 2]

Substrates: -
Products: -

?

ATP + H2O + Zn2+[side 1]

ADP + phosphate + Zn2+[side 2]

Substrates: lowest activity
Products: -

?

bilirubin/cytoplasm + ATP + H2O

bilirubin/vacuole + ADP + phosphate

-

Substrates: unconjugated substrate
Products: -

?

Cd x glutathione S2/cytoplasm + ATP + H2O

Cd x glutathione S2/vacuole + phosphate

-

Substrates: -
Products: -

?

Cd/cytoplasm + ATP + H2O

Cd/vacuole + ADP + phosphate

diazaborine/cytoplasm + ATP + H2O

diazaborine/vacuole + ADP + phosphate

-

289189

Substrates: -
Products: -

?

glutathione S-conjugated leukotriene C4/cytoplasm + ATP + H2O

glutathione S-conjugated leukotriene C4/vacuole + ADP + phosphate

-

289187, 289190

Substrates: -
Products: -

?

glutathione S-conjugates/cytoplasm + ATP + H2O

glutathione S-conjugates/vacuole + ADP + phosphate

-

Substrates: -
Products: -

?

glutathione/cytoplasm + ATP + H2O

glutathione/vacuole + ADP + phosphate

-

Substrates: -
Products: -

?

S-(2,4-dinitrophenyl)glutathione/cytoplasm + ATP + H2O

S-(2,4-dinitrophenyl)glutathione/vacuole + ADP + phosphate

-

289186, 289187, 289191

Substrates: -
Products: -

?

additional information

?

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ATP

4',4'-diisothiocyanatostilbene-2',2'-disulfonic acid

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uptake of S-(2,4-dinitrophenyl)glutathione inhibited

N-ethylmaleimide

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inhibition of phosphorylation, Cd2+ protects

654509

PAK-104P

-

42% inhibition at 0.1 mM

probenecid

-

uptake of S-(2,4-dinitrophenyl)glutathione inhibited

sulfinpyrazone

-

uptake of S-(2,4-dinitrophenyl)glutathione inhibited

vanadate

verapamil

-

IC50: 0.2 mM

vinblastine

-

IC50: 0.08 mM

additional information

-

Tus1p

-

a guanine nucleotide exchange factor, stimulates the enzyme

-

additional information

-

0.05 - 1.71

ATP

16 entries

0.0000168 - 0.0000237

bilirubin

0.187 - 0.548

estradiol-beta-17-glucuronide [side 1]

4 entries

15

glutathione

-

in vesicles

0.000758 - 0.00082

leukotriene C4 [side 1]

0.18

vanadate

Saccharomyces cerevisiae

-

IC50: 0.18 mM

0.2

verapamil

Saccharomyces cerevisiae

-

IC50: 0.2 mM

0.08

vinblastine

Saccharomyces cerevisiae

-

IC50: 0.08 mM

0.04

with Cd2+ as transport ion, at pH 7.5 and 30°C

0.06

with Co2+ as transport ion, at pH 7.5 and 30°C

0.2

with Cd2+ as transport ion, at pH 7.5 and 30°C

additional information

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measurement of growth inhibition by cadmium in wild-type and mutant strains, overview

7

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assay at

25

-

assay at

30

-

assay at

37

-

assay at

Caenorhabditis elegans

formerly named Alcaligenes eutrophus and Ralstonia metallidurans

UniProt

Drosophila melanogaster

-

Q9VF20

Uniprot

Glycine max

cultivars Westag 97 and AC Hime

749020

-

no activity in Arabidopsis thaliana ecotype Columbia

-

-

-

UniProt

Saccharomyces cerevisiae BY4741

gene YCF1

701314

-

Q0WLU3

-

intestine

Caenorhabditis elegans

-

Q0WLU3

high expression level

neuron

Caenorhabditis elegans

head neurons

-

Q0WLU3

low expression level

additional information

Q0WLU3

poor expression in roots

chloroplast

Q0WLU3

envelope membrane

9507

endoplasmic reticulum

membrane

Q0WLU3

-

P39109

-

5773

734214

additional information

Highest Expressing Human Cell Lines

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Cell Line Links

Gene Links

evolution

malfunction

physiological function

7 entries

Saccharomyces cerevisiae S288C

120000

140000

-

SDS-PAGE

160000 - 170000

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SDS-PAGE

289194

170000

171000

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nucleotide sequence

289194

185000 - 200000

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SDS-PAGE

289195

70000

Q0WLU3

x * 70000

77000

-

x * 77000, SDS-PAGE

655547

97900

Drosophila melanogaster

Q9VF20

x * 97900

additional information

4 entries

?

additional information

phosphoprotein

additional information

structures of Ycf1 mutant E1435Q at 3.4 A and 4.0 A resolution in inward-facing open conformations that capture ordered states of the intrinsically disordered regulatory domain (R-domain). R-domain phosphorylation induces a topology promoting electrostatic and hydrophobic interactions with Nucleotide Binding Domain 1 (NBD1) and the Lasso motif. These interactions are related by rigid body movements of the NBD1/R-domain complex

P39109

767586

C476A

the mutant enzyme binds 0.61 mol equiv of Co2+. The mutant enzyme shows about 4%, about 24%, and about 30% of Cd2+-, Co2+-, and Zn2+-stimulated ATP hydrolysis activity, respectively, compared to the wild type enzyme

H807A

the mutant enzyme retains the ability to bind about 1 mol equiv of all metal ions (Co2+, Zn2+, and Cd2+). The mutation results in a strong decrease in Co2+- (about 17% of wild type) and Zn2+-stimulated (about 16% of wild type) ATP hydrolysis

M254A

the mutant enzyme binds 0.34 mol and 0.26 mol equiv of Cd2+ and Co2+, respectively. The mutation dramatically reduces Cd2+- and Co2+-stimulated ATPase activity (about 8% and about 32% of wild type activity, respectively), whereas Zn2+-stimulated ATPase activity remains virtually unchanged

S474A

the mutant enzyme binds 0.45 mol equiv of Co2+. The mutation modestly decreases Co2+-stimulated ATP hydrolysis (about 66% of wild type activity) whereas Cd2+- and Zn2+-stimulated ATP hydrolysis remain virtually unchanged

S474A/C476A

the mutant enzyme shows about 2%, about 16%, and about 20% of Cd2+-, Co2+-, and Zn2+-stimulated ATP hydrolysis activity, respectively, compared to the wild type enzyme

D398A

-

non-phosphorylatable protein, no stimulation by Cd2+ but rather inhibition at high Cd2+ concentrations

655547

A910G

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mutants more sensitive to Cd2+ than wild type strain, higher activity for glutathione S-conjugated leukotriene C4

D777N

D777N/A1003V

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intragenic suppressor of mutation D777N

D777N/A1021T

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intragenic suppressor of mutation D777N, functionally important residue

D777N/A1021V

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intragenic suppressor of mutation D777N, functionally important residue

D777N/F565L

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intragenic suppressor of mutation D777N, involved in determination of substrate specificity

D777N/G1207D

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intragenic suppressor of mutation D777N, functionally important residue

D777N/G1207S

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intragenic suppressor of mutation D777N, functionally important residue

D777N/N1027D

-

intragenic suppressor of mutation D777N

D777N/Q1107R

-

intragenic suppressor of mutation D777N, involved in determination of substrate specificity

D777N/R1415G

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intragenic suppressor of mutation D777N, functionally important residue

D777N/S1212L

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intragenic suppressor of mutation D777N, functionally important residue

D777N/S674L

-

intragenic suppressor of mutation D777N

D777N/V543I

-

intragenic suppressor of mutation D777N, involved in determination of substrate specificity

D777N/W1225C

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intragenic suppressor of mutation D777N, involved in determination of substrate specificity

D821G

-

mutants with Ycf1p in vivo activity as wild type strain

E1435Q

P39109

NBD2 walker B mutant, substantially lowered ATPase activity

767586

E709Q

-

mutants with Ycf1p in vivo activity as wild type strain

E927K

-

mutants exhibit a Cd2+ hypersensitive phenotype

G1306E

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mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4

G1311R

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mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4

G1413D

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mutants exhibit a Cd2+ hypersensitive phenotype

G663V

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mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4

G756D

G756S

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substitution mutation, unable to complement the cadmium hypersensitivity of the introduced strain

G835R

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mutants more sensitive to Cd2+ than wild type strain, partially reduced transport activity for glutathione S-conjugated leukotriene C4

I711S

-

mutants exhibit a Cd2+ hypersensitive phenotype

I840P

-

mutants more sensitive to Cd2+ than wild type strain, partially reduced transport activity for glutathione S-conjugated leukotriene C4

K669M

L817S

-

mutants more sensitive to Cd2+ than wild type strain

L825T

-

mutants more sensitive to Cd2+ than wild type strain

L826S

-

mutants more sensitive to Cd2+ than wild type strain, partially reduced transport activity for glutathione S-conjugated leukotriene C4

N1366K

-

mutants exhibit a Cd2+ hypersensitive phenotype, partially reduced transport activity for glutathione S-conjugated leukotriene C4

Q1148P

-

mutants more sensitive to Cd2+ than wild type strain

R1143C

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mutants more sensitive to Cd2+ than wild type strain, partially reduced transport activity for glutathione S-conjugated leukotriene C4

S251A

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the site-directed mutation inhibits phosphorylation, the mutant exhibits increased resistance to cadmium in vivo and increased Ycf1p-dependent transport of [3H]estradiol-beta-17-glucuronide in vitro as compared with wild-type Ycf1p

S251E

-

the site-directed mutation mimics phosphorylation, activity is restored to the wild-type level for mutant Ycf1-S251E

S908A

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replacement of Ser908 by alanine

S908A/T911A

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the mutant shows a 50% diminished expression of Ycf1p compared to the wild-type enzyme

S908D

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replacement of Ser908 by aspartate

S908E

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replacement of Ser908 by glutamate

S908T

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replacement of Ser908 by threonine

Y855L

-

mutants more sensitive to Cd2+ than wild type strain, higher activity for glutathione S-conjugated leukotriene C4

additional information

15 entries

Pca1 is a short-lived proteinwith t1?2 below 5 min and is subject to ubiquitination when cells are growing in media lacking cadmium

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698831

-85°C, vacuolar membrane preparations

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Ni-charged HiTrap column chromatography and Ni-NTA column chromatography

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construction of an integrated membrane yeast two-hybrid, iMYTH, screen and identification of two kinase genes, CKA1 and HAL5, deletion of which increases Ycf1p function

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expressed in Escherichia coli BL21(DE3) cells

expressed in Oryza sativa mesophyll protoplasts

Glycine max

-

749020

expressed in Sf21 insect cells

-

expression in Schizosaccharomyces pombe

Drosophila melanogaster

Q9VF20

gene YCF1, semiquantitative expression analysis in absence and presence of Cd2+, overview

-

701314

OSA1 expression analysis, and phylogenetic analysis, overview. No complementation of Saccharomyces cerevisiae mutant strain W303-1A abc1THIS3 by the wild-type enzyme

Q0WLU3

Pca1 expression analysis

-

698831

phylogenetic analysis

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the pYES2 plasmid is used for overexpression of EGFP-N2, YCF1, or YCF-EGFP-N2 in different Saccharomyces cerevisiae strains

-

685407

transgenic expression of wild-type and mutant enzymes in Arabidopsis thaliana Col-0 ecotype, which is sensitive to Cd. EGFP-tagged mutant OsMA3-C788 is bombarded into onion epidermal cell, the green fluorescence of the OsHMA3-C788 fusion protein is observed as an outline of the cell shape and is also delineated the nucleus

Oryza sativa

Q8H384

733874

endoplasmic reticulum-associated degradation (ERAD) system is responsible for the regulation of Pca1 expression by cadmium. Direct cadmium sensing at the endoplasmic reticulum by a degron in Pca1 leads to an escape of Pca1 from endoplasmic reticulum-associated degradation

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713395

in the absence of its cadmium substrate, Pca1 is rapidly degraded through the endoplasmic reticulum-associated degradation pathway

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713395

the expression of gene YFC1 is downregulated in presence of Cd2+, which affects the YFC1 promoter

agriculture

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improvement of heavy metal, salt or xenobiotic chemical tolerance in plants

670617

environmental protection