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Escherichia coli K-12 substr. MG1655 Transporter: phosphate ABC transporter

Synonyms: pstSCAB


Subunit composition of phosphate ABC transporter = [PstB]2[PstA][PstC][PstS]
         phosphate ABC transporter - ATP binding subunit = PstB (summary available)
         phosphate ABC transporter - membrane subunit = PstA (summary available)
         phosphate ABC transporter - membrane subunit = PstC (summary available)
         phosphate ABC transporter - periplasmic binding protein = PstS (extended summary available)

Summary:
PstSCAB is an ATP-dependent phosphate uptake system that is a member of the ATP-Binding Cassette (ABC) Superfamily of transporters [Wu95]. PstSCAB is responsible for inorganic phosphate (Pi) uptake under Pi starvation conditions. Inorganic phosphate is an essential component in cellular function since phosphorylation of nucleic acids, lipids, sugars, and proteins are important for gene regulation and signaling [Torriani90]. Based on sequence similarity, PstA and PstC are the membrane components of the ABC transporter, while PstS is the periplasmic phosphate binding protein [Torriani90], and PstB is the ATP-binding component of the ABC transporter [Torriani90]. Whole cell transport assay indicates that the Pst system has a Km of 0.20 μM [Cox81]. Transcription of the Pst system is induced by Pi starvation, as opposed to the Pit phosphate transport system that is expressed regardless of Pi level [Torriani90].

The role of the Pst system in arsenate transport has been studied. A strain dependent on the Pst system for phosphate transport grows at a reduced rate in the presence of arsenate. This strain does not accumulate radioactive arsenate and shows only a transient decrease in intracellular ATP levels after addition of arsenate to the culture media [Willsky80].

Gene-Reaction Schematic

Gene-Reaction Schematic


Enzymatic reaction of: transport of phosphate (phosphate ABC transporter)

Inferred from experimentInferred by computational analysis

EC Number: 3.6.3.27

Transport reaction diagram for transport of phosphate

Inhibitors (Competitive): arsenate (Kic = 23µM) [Willsky80]Kinetic Parameters:
Substrate Km (μM) Citations
ATP 23.9 [Sarin01, BRENDA14]

Subunit of phosphate ABC transporter: phosphate ABC transporter - ATP binding subunit

Synonyms: PhoT

Gene: pstB Accession Numbers: EG10783 (EcoCyc), b3725, ECK3718

Locations: inner membrane, cytosol

Sequence Length: 257 AAs

Molecular Weight: 29.027 kD (from nucleotide sequence)


GO Terms:
Biological Process:
Inferred from experimentInferred by computational analysisGO:0006817 - phosphate ion transport [UniProtGOA11a, GOA06, GOA01a, Cox81]
Inferred by computational analysisGO:0006810 - transport [UniProtGOA11a]
Inferred by computational analysisGO:0008152 - metabolic process [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0035435 - phosphate ion transmembrane transport [GOA01a]
Molecular Function:
Inferred from experimentInferred by computational analysisGO:0015415 - phosphate ion transmembrane-transporting ATPase activity [GOA06, GOA01, GOA01a, Cox81]
Inferred by computational analysisGO:0000166 - nucleotide binding [UniProtGOA11a]
Inferred by computational analysisGO:0005315 - inorganic phosphate transmembrane transporter activity [GOA01a]
Inferred by computational analysisGO:0005524 - ATP binding [UniProtGOA11a, GOA06, GOA01a]
Inferred by computational analysisGO:0016787 - hydrolase activity [UniProtGOA11a]
Inferred by computational analysisGO:0016887 - ATPase activity [GOA01a]
Cellular Component:
Inferred from experimentInferred by computational analysisGO:0005886 - plasma membrane [UniProtGOA11, UniProtGOA11a, DiazMejia09, Zhang07]
GO:0005829 - cytosol []
Inferred by computational analysisGO:0016020 - membrane [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0043190 - ATP-binding cassette (ABC) transporter complex [GOA06]

MultiFun Terms: metabolismmetabolism of other compoundsphosphorous metabolism
transportChannel-type TransportersPyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active TransportersThe ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake PermeasesABC superfamily ATP binding cytoplasmic component

Unification Links: DIP:DIP-47851N, EcoliWiki:b3725, Mint:MINT-1233567, PR:PRO_000023622, Pride:P0AAH0, Protein Model Portal:P0AAH0, RefSeq:NP_418181, SMR:P0AAH0, UniProt:P0AAH0

Relationship Links: InterPro:IN-FAMILY:IPR003439, InterPro:IN-FAMILY:IPR003593, InterPro:IN-FAMILY:IPR005670, InterPro:IN-FAMILY:IPR015850, InterPro:IN-FAMILY:IPR017871, InterPro:IN-FAMILY:IPR027417, Pfam:IN-FAMILY:PF00005, Prosite:IN-FAMILY:PS00211, Prosite:IN-FAMILY:PS50893, Prosite:IN-FAMILY:PS51238, Smart:IN-FAMILY:SM00382

Summary:
pstB is one of a network of genes believed to play a role in promoting the stress-induced mutagenesis (SIM) response of E. coli K-12 [Al12].

Essentiality data for pstB knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enrichedYes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB LennoxYes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Subunit of phosphate ABC transporter: phosphate ABC transporter - membrane subunit

Synonyms: PhoT

Gene: pstA Accession Numbers: EG10782 (EcoCyc), b3726, ECK3719

Locations: inner membrane

Sequence Length: 296 AAs

Molecular Weight: 32.322 kD (from nucleotide sequence)


GO Terms:
Biological Process:
Inferred from experimentInferred by computational analysisGO:0006817 - phosphate ion transport [UniProtGOA11a, Cox81]
Inferred from experimentGO:0006974 - cellular response to DNA damage stimulus [Khil02]
Inferred by computational analysisGO:0006810 - transport [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0035435 - phosphate ion transmembrane transport [Gaudet10, GOA01a]
Molecular Function:
Inferred from experimentGO:0015415 - phosphate ion transmembrane-transporting ATPase activity [Cox81]
Inferred by computational analysisGO:0005315 - inorganic phosphate transmembrane transporter activity [GOA01a]
Cellular Component:
Inferred from experimentInferred by computational analysisGO:0005887 - integral component of plasma membrane [GOA01a, Drew02]
Inferred by computational analysisGO:0005886 - plasma membrane [UniProtGOA11, UniProtGOA11a, DiazMejia09]
Inferred by computational analysisGO:0016020 - membrane [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0016021 - integral component of membrane [UniProtGOA11a]

MultiFun Terms: cell structuremembrane
metabolismmetabolism of other compoundsphosphorous metabolism
transportChannel-type TransportersPyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active TransportersThe ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake PermeasesABC superfamily, membrane component

Unification Links: EcoliWiki:b3726, PR:PRO_000023621, Protein Model Portal:P07654, RefSeq:NP_418182, UniProt:P07654

Relationship Links: InterPro:IN-FAMILY:IPR000515, InterPro:IN-FAMILY:IPR005672, Pfam:IN-FAMILY:PF00528, Prosite:IN-FAMILY:PS50928

Summary:
Protein topology in the inner membrane has been determined [Drew02].

Essentiality data for pstA knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enrichedYes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB LennoxYes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Subunit of phosphate ABC transporter: phosphate ABC transporter - membrane subunit

Synonyms: PhoW

Gene: pstC Accession Numbers: EG10784 (EcoCyc), b3727, ECK3720

Locations: inner membrane

Sequence Length: 319 AAs

Molecular Weight: 34.121 kD (from nucleotide sequence)


GO Terms:
Biological Process:
Inferred from experimentInferred by computational analysisGO:0006817 - phosphate ion transport [UniProtGOA11a, GOA01a, Cox81]
Inferred from experimentGO:0035435 - phosphate ion transmembrane transport [Cox81]
Inferred by computational analysisGO:0006810 - transport [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0055085 - transmembrane transport [GOA01a]
Molecular Function:
Inferred from experimentGO:0015415 - phosphate ion transmembrane-transporting ATPase activity [Cox81]
Inferred by computational analysisGO:0005315 - inorganic phosphate transmembrane transporter activity [GOA01a]
Cellular Component:
Inferred from experimentInferred by computational analysisGO:0005886 - plasma membrane [UniProtGOA11, UniProtGOA11a, DiazMejia09, Daley05]
Inferred by computational analysisGO:0005887 - integral component of plasma membrane [Gaudet10]
Inferred by computational analysisGO:0016020 - membrane [UniProtGOA11a, GOA01a]
Inferred by computational analysisGO:0016021 - integral component of membrane [UniProtGOA11a, GOA01a]

MultiFun Terms: cell structuremembrane
metabolismmetabolism of other compoundsphosphorous metabolism
transportChannel-type TransportersPyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active TransportersThe ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake PermeasesABC superfamily, membrane component

Unification Links: EcoliWiki:b3727, PR:PRO_000023623, Protein Model Portal:P0AGH8, RefSeq:NP_418183, UniProt:P0AGH8

Relationship Links: InterPro:IN-FAMILY:IPR000515, InterPro:IN-FAMILY:IPR011864, Pfam:IN-FAMILY:PF00528, Prosite:IN-FAMILY:PS50928

Summary:
pstC is one of a network of genes believed to play a role in promoting the stress-induced mutagenesis (SIM) response of E. coli K-12 [Al12].

Essentiality data for pstC knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enrichedYes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB LennoxYes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Subunit of phosphate ABC transporter: phosphate ABC transporter - periplasmic binding protein

Synonyms: PhoS, NmpA

Gene: pstS Accession Numbers: EG10734 (EcoCyc), b3728, ECK3721

Locations: inner membrane, periplasmic space

Sequence Length: 346 AAs

Molecular Weight: 37.024 kD (from nucleotide sequence)


GO Terms:
Biological Process:
Inferred from experimentGO:0006974 - cellular response to DNA damage stimulus [Khil02]
Inferred by computational analysisGO:0006810 - transport [UniProtGOA11a]
Inferred by computational analysisGO:0006817 - phosphate ion transport [UniProtGOA11a, Gaudet10]
Inferred by computational analysisGO:0016036 - cellular response to phosphate starvation [Gaudet10]
Inferred by computational analysisGO:0035435 - phosphate ion transmembrane transport [GOA01a]
Molecular Function:
Inferred from experimentGO:0015415 - phosphate ion transmembrane-transporting ATPase activity [Cox81]
Inferred by computational analysisGO:0042301 - phosphate ion binding [Gaudet10, GOA01a]
Cellular Component:
Inferred by computational analysisGO:0005886 - plasma membrane [Gaudet10]
GO:0030288 - outer membrane-bounded periplasmic space []
Inferred by computational analysisGO:0042597 - periplasmic space [UniProtGOA11, UniProtGOA11a]
Inferred by computational analysisGO:0043190 - ATP-binding cassette (ABC) transporter complex [GOA01a]

MultiFun Terms: metabolismmetabolism of other compoundsphosphorous metabolism
transportChannel-type TransportersPyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active TransportersThe ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake PermeasesABC superfamily, periplasmic binding component

Unification Links: DIP:DIP-48241N, EcoliWiki:b3728, Mint:MINT-1224444, ModBase:P0AG82, PR:PRO_000023624, Pride:P0AG82, Protein Model Portal:P0AG82, RefSeq:NP_418184, SMR:P0AG82, Swiss-Model:P0AG82, UniProt:P0AG82

Relationship Links: InterPro:IN-FAMILY:IPR005673, InterPro:IN-FAMILY:IPR024370, PDB:Structure:1A40, PDB:Structure:1A54, PDB:Structure:1A55, PDB:Structure:1IXG, PDB:Structure:1IXH, PDB:Structure:1IXI, PDB:Structure:1OIB, PDB:Structure:1PBP, PDB:Structure:1QUI, PDB:Structure:1QUJ, PDB:Structure:1QUK, PDB:Structure:1QUL, PDB:Structure:2ABH, Pfam:IN-FAMILY:PF12849

Summary:
PstS is the periplasmic, phosphate binding protein of the high affinity phosphate transport sytem in E. coli K-12.

PstS consists of two globular domains which form a phosphate binding cleft [Luecke90]. Phosphate is held in place by 12 hydrogen bonds [Luecke90]. PstS binds monobasic (H2PO4-) and dibasic (H2PO42-) phosphates [Wang94].

PstS can discriminate phosphate from arsenate up to an 800 fold molar excess of arsenate [Elias12]. A pstS D81N mutant has no effect on phosphate binding but disturbs the discrimination against arsenate approximately 10-fold and the discrimination against sulfate about 100-fold [Elias12]. The asparagine residue at position 81 forms a very short hydrogen bond with phosphate [Wang97].

pstS insertion mutants were identified in a genetic screen for genes that are important for survival of exposure to ionizing radiation (IR). A pstS deletion mutant has a substantial decrease in IR survival [Byrne14].

Essentiality data for pstS knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enrichedYes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB LennoxYes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

References

Al12: Al Mamun AA, Lombardo MJ, Shee C, Lisewski AM, Gonzalez C, Lin D, Nehring RB, Saint-Ruf C, Gibson JL, Frisch RL, Lichtarge O, Hastings PJ, Rosenberg SM (2012). "Identity and function of a large gene network underlying mutagenic repair of DNA breaks." Science 338(6112);1344-8. PMID: 23224554

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

BRENDA14: BRENDA team (2014). Imported from BRENDA version existing on Aug 2014.

Byrne14: Byrne RT, Chen SH, Wood EA, Cabot EL, Cox MM (2014). "Escherichia coli genes and pathways involved in surviving extreme exposure to ionizing radiation." J Bacteriol 196(20);3534-45. PMID: 25049088

Cox81: Cox GB, Rosenberg H, Downie JA, Silver S (1981). "Genetic analysis of mutants affected in the Pst inorganic phosphate transport system." J Bacteriol 1981;148(1);1-9. PMID: 7026529

Daley05: Daley DO, Rapp M, Granseth E, Melen K, Drew D, von Heijne G (2005). "Global topology analysis of the Escherichia coli inner membrane proteome." Science 308(5726);1321-3. PMID: 15919996

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Drew02: Drew D, Sjostrand D, Nilsson J, Urbig T, Chin CN, de Gier JW, von Heijne G (2002). "Rapid topology mapping of Escherichia coli inner-membrane proteins by prediction and PhoA/GFP fusion analysis." Proc Natl Acad Sci U S A 99(5);2690-5. PMID: 11867724

Elias12: Elias M, Wellner A, Goldin-Azulay K, Chabriere E, Vorholt JA, Erb TJ, Tawfik DS (2012). "The molecular basis of phosphate discrimination in arsenate-rich environments." Nature 491(7422);134-7. PMID: 23034649

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gaudet10: Gaudet P, Livstone M, Thomas P (2010). "Annotation inferences using phylogenetic trees." PMID: 19578431

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Khil02: Khil PP, Camerini-Otero RD (2002). "Over 1000 genes are involved in the DNA damage response of Escherichia coli." Mol Microbiol 44(1);89-105. PMID: 11967071

Luecke90: Luecke H, Quiocho FA (1990). "High specificity of a phosphate transport protein determined by hydrogen bonds." Nature 347(6291);402-6. PMID: 2215649

Sarin01: Sarin J, Aggarwal S, Chaba R, Varshney GC, Chakraborti PK (2001). "B-subunit of phosphate-specific transporter from Mycobacterium tuberculosis is a thermostable ATPase." J Biol Chem 276(48);44590-7. PMID: 11567022

Torriani90: Torriani A (1990). "From cell membrane to nucleotides: the phosphate regulon in Escherichia coli." Bioessays 1990;12(8);371-6. PMID: 2241934

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Wang94: Wang Z, Choudhary A, Ledvina PS, Quiocho FA (1994). "Fine tuning the specificity of the periplasmic phosphate transport receptor. Site-directed mutagenesis, ligand binding, and crystallographic studies." J Biol Chem 269(40);25091-4. PMID: 7929197

Wang97: Wang Z, Luecke H, Yao N, Quiocho FA (1997). "A low energy short hydrogen bond in very high resolution structures of protein receptor--phosphate complexes." Nat Struct Biol 4(7);519-22. PMID: 9228942

Willsky80: Willsky GR, Malamy MH (1980). "Effect of arsenate on inorganic phosphate transport in Escherichia coli." J Bacteriol 144(1);366-74. PMID: 6998959

Wu95: Wu LF, Mandrand-Berthelot MA (1995). "A family of homologous substrate-binding proteins with a broad range of substrate specificity and dissimilar biological functions." Biochimie 1995;77(9);744-50. PMID: 8789466

Zhang07: Zhang N, Chen R, Young N, Wishart D, Winter P, Weiner JH, Li L (2007). "Comparison of SDS- and methanol-assisted protein solubilization and digestion methods for Escherichia coli membrane proteome analysis by 2-D LC-MS/MS." Proteomics 7(4);484-93. PMID: 17309111


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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