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Information on EC 7.2.2.2 - ABC-type Cd2+ transporter and Organism(s) Saccharomyces cerevisiae and UniProt Accession P39109

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IUBMB Comments
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.
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Saccharomyces cerevisiae
UNIPROT: P39109
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Word Map
The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
ycf1p, oshma3, yor1p, hmt-1, yeast cadmium factor, yeast cadmium factor 1, cadmium-transporting atpase, hmt1 protein, heavy metal-associated atpase, heavy metal tolerance factor 1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ATP phosphohydrolase (heavy-metal-exporting)
-
-
-
-
cadmium-transporting ATPase
cadmium-transporting P1B-type ATPase
-
-
yeast cadmium factor
-
-
-
-
yeast cadmium factor 1
-
-
additional information
-
the enzyme belongs to the ABCC subfamily of ABC transporters
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
transmembrane transport
SYSTEMATIC NAME
IUBMB Comments
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.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + H2O + Cd2+/in
ADP + phosphate + Cd2+/out
show the reaction diagram
ATP + H2O + estradiol-beta-17-glucuronide [side 1]
ADP + phosphate + estradiol-beta-17-glucuronide [side 2]
show the reaction diagram
-
-
-
-
?
ATP + H2O + leukotriene C4 [side 1]
ADP + phosphate + leukotriene C4 [side 2]
show the reaction diagram
-
-
-
-
?
bilirubin/cytoplasm + ATP + H2O
bilirubin/vacuole + ADP + phosphate
show the reaction diagram
-
unconjugated substrate
-
-
?
Cd x glutathione S2/cytoplasm + ATP + H2O
Cd x glutathione S2/vacuole + phosphate
show the reaction diagram
-
-
-
-
?
Cd/cytoplasm + ATP + H2O
Cd/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
Cd2+/cytoplasm + ATP + H2O
Cd2+/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
diazaborine/cytoplasm + ATP + H2O
diazaborine/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione S-conjugated leukotriene C4/cytoplasm + ATP + H2O
glutathione S-conjugated leukotriene C4/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione S-conjugates/cytoplasm + ATP + H2O
glutathione S-conjugates/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione/cytoplasm + ATP + H2O
glutathione/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
S-(2,4-dinitrophenyl)glutathione/cytoplasm + ATP + H2O
S-(2,4-dinitrophenyl)glutathione/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + H2O + Cd2+/in
ADP + phosphate + Cd2+/out
show the reaction diagram
bilirubin/cytoplasm + ATP + H2O
bilirubin/vacuole + ADP + phosphate
show the reaction diagram
-
unconjugated substrate
-
-
?
Cd x glutathione S2/cytoplasm + ATP + H2O
Cd x glutathione S2/vacuole + phosphate
show the reaction diagram
-
-
-
-
?
Cd/cytoplasm + ATP + H2O
Cd/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
diazaborine/cytoplasm + ATP + H2O
diazaborine/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione S-conjugated leukotriene C4/cytoplasm + ATP + H2O
glutathione S-conjugated leukotriene C4/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione S-conjugates/cytoplasm + ATP + H2O
glutathione S-conjugates/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione/cytoplasm + ATP + H2O
glutathione/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
S-(2,4-dinitrophenyl)glutathione/cytoplasm + ATP + H2O
S-(2,4-dinitrophenyl)glutathione/vacuole + ADP + phosphate
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4',4'-diisothiocyanatostilbene-2',2'-disulfonic acid
-
uptake of S-(2,4-dinitrophenyl)glutathione inhibited
PAK-104P
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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
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Tus1p
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a guanine nucleotide exchange factor, stimulates the enzyme
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 1.71
ATP
0.0000168 - 0.0000237
bilirubin
0.187 - 0.548
estradiol-beta-17-glucuronide [side 1]
15
glutathione
-
in vesicles
0.000758 - 0.00082
leukotriene C4 [side 1]
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
measurement of growth inhibition by cadmium in wild-type and mutant strains, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
the N terminus of Pca1 contains a targeting signal for ERAD. Degradation of Pca1 and cadmium sensing occur at the endoplasmic reticulum
Manually annotated by BRENDA team
additional information
-
Pca1 contains a cytosolic cysteine-rich N-terminal extension of nearly 400 residues
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme belongs to the ATP-binding cassette class C (ABCC) transporter family
malfunction
deletion of YCF1 attenuates in vitro vacuole fusion by up to 40% relative to wild-type vacuoles, plasmid-expressed wild-type Ycf1p rescues the deletion phenotype. Ycf1p containing a mutation of the conserved Lys669 to Met in the Walker A box of the first nucleotide-binding domain (Ycf1pK669M) is unable to complement the fusion defect of DELTAycf1 vacuoles. Also, deleting YCF1 causes a striking decrease in vacuolar levels of the soluble SNARE Vam7p, whereas total cellular levels are not altered. The attenuated fusion of DELTAycf1 vacuoles is rescued by the addition of recombinant Vam7p to in vitro experiments, exogenous Vam7p restores Ca2+ efflux in DELTAycf1 vacuoles. Phenotypes, overview
physiological function
the enzyme Ycf1p is a Cd2+ transporter, it regulates the transport of cadmium, mercury, and other toxins into the vacuole lumen to detoxify the cytoplasm. The ATPase activity of Ycf1p is required for its function in regulating fusion. The enzyme physically interacts with a wide array of proteins, including factors that regulate vacuole homeostasis. Role of Ycf1p and other ABCC transporters in the regulation of vacuole homotypic fusion, overview. Ycf1p contributes in the recruitment of Vam7p to the vacuole for efficient membrane fusion
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
120000
140000
-
SDS-PAGE
160000 - 170000
-
SDS-PAGE
170000
171000
-
nucleotide sequence
additional information
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Pca1 possesses a metal-responding degradation signal encompassing amino acids 250-350, which functions autonomously in a metal-responsive manner, being part of the cytosolic cysteine-rich N-terminal extension of nearly 400 residues
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
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the enzyme is activated by phosphorylation in its ABC core domain at residues Ser908 and Thr911, while phosphorylation of Ser251 in the Ycf1p N-terminal extension negatively regulates activity
additional information
-
cadmium sensing by cysteine residues within the domain, the N-terminal metal-responding degradation signal encompassing amino acids 250-350, circumvents ubiquitination and degradation of Pca1
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K669M
enzyme Ycf1p containing a mutation of the conserved Lys669 to Met in the Walker A box of the first nucleotide-binding domain (Ycf1pK669M) is unable to complement the fusion defect of DELTAycf1 vacuoles
A910G
-
mutants more sensitive to Cd2+ than wild type strain, higher activity for glutathione S-conjugated leukotriene C4
D777N
D777N/A1003V
-
intragenic suppressor of mutation D777N
D777N/A1021T
-
intragenic suppressor of mutation D777N, functionally important residue
D777N/A1021V
-
intragenic suppressor of mutation D777N, functionally important residue
D777N/F565L
-
intragenic suppressor of mutation D777N, involved in determination of substrate specificity
D777N/G1207D
-
intragenic suppressor of mutation D777N, functionally important residue
D777N/G1207S
-
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
-
intragenic suppressor of mutation D777N, functionally important residue
D777N/S1212L
-
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
-
intragenic suppressor of mutation D777N, involved in determination of substrate specificity
D821G
-
mutants with Ycf1p in vivo activity as wild type strain
E709Q
-
mutants with Ycf1p in vivo activity as wild type strain
E927K
-
mutants exhibit a Cd2+ hypersensitive phenotype
G1306E
-
mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4
G1311R
-
mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4
G1413D
-
mutants exhibit a Cd2+ hypersensitive phenotype
G663V
-
mutants exhibit a Cd2+ hypersensitive phenotype, abolished transport activity for glutathione S-conjugated leukotriene C4
G756D
G756S
-
substitution mutation, unable to complement the cadmium hypersensitivity of the introduced strain
G835R
-
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
-
substitution mutation, unable to complement the cadmium hypersensitivity of the introduced strain
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
-
mutants more sensitive to Cd2+ than wild type strain, partially reduced transport activity for glutathione S-conjugated leukotriene C4
S251A
-
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
-
replacement of Ser908 by alanine
S908A/T911A
-
the mutant shows a 50% diminished expression of Ycf1p compared to the wild-type enzyme
S908D
-
replacement of Ser908 by aspartate
S908E
-
replacement of Ser908 by glutamate
S908T
-
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
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Pca1 is a short-lived proteinwith t1?2 below 5 min and is subject to ubiquitination when cells are growing in media lacking cadmium
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-85°C, vacuolar membrane preparations
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
-
expressed in Sf21 insect cells
-
gene YCF1, semiquantitative expression analysis in absence and presence of Cd2+, overview
-
Pca1 expression analysis
-
the pYES2 plasmid is used for overexpression of EGFP-N2, YCF1, or YCF-EGFP-N2 in different Saccharomyces cerevisiae strains
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
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
-
in the absence of its cadmium substrate, Pca1 is rapidly degraded through the endoplasmic reticulum-associated degradation pathway
-
the expression of gene YFC1 is downregulated in presence of Cd2+, which affects the YFC1 promoter
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
environmental protection
-
overexpression of this fungal transporter in plants can be useful for phytoremediation of lead and cadmium polluted soils
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Li, Z.S.; Szczypka, M.; Lu, Y.P.; Thiele ,D.J.; Rea, P.A.
The yeast cadmium factor protein (YCF1) is a vacuolar glutathione S-conjugate pump
J. Biol. Chem.
217
6509-6517
1996
Saccharomyces cerevisiae
-
Manually annotated by BRENDA team
Ren, X.Q.; Furukawa, T.; Chen, Z.S.; Okumura, H.; Aoki, S.; Sumizawa, T.; Tani, A.; Komatsu, M.; Mei, X.D.; Akiyama S.
Functional comparison between YCF1 and MRP1 expresses in Sf21 insect cells
Biochem. Biophys. Res. Commun.
270
608-615
2000
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Petrovic, S.; Pascolo, L.; Gallo, R.; Cupelli, F.; Ostrow, J.D.; Goffeau, A.; Tiribelli, C.; Bruschi, C.V.
The products of YCF1 and YLL015w (BPT1) cooperate for the ATP-dependent vacuolar transport of unconjugated bilirubin in Saccharomyces cerevisiae
Yeast
16
561-571
2000
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Jungwirth, H.; Wendler, F.; Platzer, B.; Bergler, H.; Hgenauer, G.
Diazaborine resistance in yeast involves the efflux pumps Ycf1p and Flr1p and is enhanced by a gain-of-function allele of gene YAP1
Eur. J. Biochem.
267
4809-4826
2000
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Falcon-Perez, J.M.; Mazon, M.J.; Molano, J.; Eraso, P.
Functional domain analysis of the yeast ABC transporter Ycf1p by site-directed mutagenesis
J. Biol. Chem.
274
23584-23590
1999
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Rebbeor, J.F.; Connolly, G.C.; Dumont, M.E.; Ballatori, N.
ATP-dependent transport of reduced glutathione on YCF1, the yeast orthologue of mammalian multidrug resistance associated proteins
J. Biol. Chem.
273
33449-33454
1998
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Wemmie, J.A.; Moye-Rowley, W.S.
Mutational analysis of the Saccharomyces cerevisiae ATP-binding cassette transporter protein Ycf1p
Mol. Microbiol.
25
683-694
1997
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Wemmie, J.A.; Szczypka, M.S.; Thiele, D.J.; Moye-Rowley, W.S.
Cadmium tolerance mediated by the yeast AP-1 protein requires the presence of an ATP-binding cassette transporter-encoding gene, YCF1
J. Biol. Chem.
269
32592-32597
1994
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Szczypka, M.S.; Wemmie, J.A.; Moye-Rowley, W.S.; Thiele, D.J.
A yeast metal resistance protein similar to human cystic fibrosis transmembrane conductance regulator (CFTR) and multidrug resistance-associated protein
J. Biol. Chem.
269
22853-22857
1994
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Falcon-Perez, J.M.; Martinez-Burgos, M.; Molano, J.; Mazon, M.J.; Eraso, P.
Domain interactions in the yeast ATP binding cassette transporter Ycf1p: intragenic suppressor analysis of mutations in the nucleotide binding domains
J. Bacteriol.
183
4761-4770
2001
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Eraso, P.; Martinez-Burgos, M.; Falcon-Perez, J.M.; Portillo, F.; Mazon, M.J.
Ycf1-dependent cadmium detoxification by yeast requires phosphorylation of residues Ser908 and Thr911
FEBS Lett.
577
322-326
2004
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Preveral, S.; Ansoborlo, E.; Mari, S.; Vavasseur, A.; Forestier, C.
Metal(loid)s and radionuclides cytotoxicity in Saccharomyces cerevisiae. Role of YCF1, glutathione and effect of buthionine sulfoximine
Biochimie
88
1651-1663
2006
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Nagy, Z.; Montigny, C.; Leverrier, P.; Yeh, S.; Goffeau, A.; Garrigos, M.; Falson, P.
Role of the yeast ABC transporter Yor1p in cadmium detoxification
Biochimie
88
1665-1671
2006
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Paumi, C.M.; Chuk, M.; Chevelev, I.; Stagljar, I.; Michaelis, S.
Negative regulation of the yeast ABC transporter Ycf1p by phosphorylation within its N-terminal extension
J. Biol. Chem.
283
27079-27088
2008
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Adle, D.J.; Lee, J.
Expressional control of a cadmium-transporting P1B-type ATPase by a metal sensing degradation signal
J. Biol. Chem.
283
31460-31468
2008
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Mielniczki-Pereira, A.A.; Schuch, A.Z.; Bonatto, D.; Cavalcante, C.F.; Vaitsman, D.S.; Riger, C.J.; Eleutherio, E.C.; Henriques, J.A.
The role of the yeast ATP-binding cassette Ycf1p in glutathione and cadmium ion homeostasis during respiratory metabolism
Toxicol. Lett.
180
21-27
2008
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4741
Manually annotated by BRENDA team
Adle, D.J.; Wei, W.; Smith, N.; Bies, J.J.; Lee, J.
Cadmium-mediated rescue from ER-associated degradation induces expression of its exporter
Proc. Natl. Acad. Sci. USA
106
10189-10194
2009
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Sasser, T.L.; Lawrence, G.; Karunakaran, S.; Brown, C.; Fratti, R.A.
The yeast ATP-binding cassette (ABC) transporter Ycf1p enhances the recruitment of the soluble SNARE Vam7p to vacuoles for efficient membrane fusion
J. Biol. Chem.
288
18300-18310
2013
Saccharomyces cerevisiae (P39109), Saccharomyces cerevisiae
Manually annotated by BRENDA team