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Escherichia coli K-12 substr. MG1655 Polypeptide: phosphate ABC transporter - periplasmic binding protein



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

Synonyms: R2pho, T, nmpA, phoR2a, phoS

Regulation Summary Diagram: ?

Component of: phosphate ABC transporter (extended summary available)

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 [Wang94a].

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 [Wang97g].

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].

Gene Citations: [Magota84, Amemura85, Steed93, Surin85, Aguena02]

Locations: inner membrane, periplasmic space

Map Position: [3,908,508 <- 3,909,548] (84.24 centisomes)
Length: 1041 bp / 346 aa

Molecular Weight of Polypeptide: 37.024 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0012190 , CGSC:17998 , DIP:DIP-48241N , EchoBASE:EB0727 , EcoGene:EG10734 , EcoliWiki:b3728 , Mint:MINT-1224444 , ModBase:P0AG82 , OU-Microarray:b3728 , PortEco:pstS , PR:PRO_000023624 , Pride:P0AG82 , Protein Model Portal:P0AG82 , RefSeq:NP_418184 , RegulonDB:EG10734 , SMR:P0AG82 , String:511145.b3728 , 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

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006974 - cellular response to DNA damage stimulus Inferred from experiment [Khil02]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11]
GO:0006817 - phosphate ion transport Inferred by computational analysis [UniProtGOA11]
GO:0006950 - response to stress Inferred by computational analysis [UniProtGOA11]
GO:0035435 - phosphate ion transmembrane transport Inferred by computational analysis [GOA01a]
Molecular Function: GO:0015415 - phosphate ion transmembrane-transporting ATPase activity Inferred from experiment [Cox81a]
GO:0042301 - phosphate ion binding Inferred by computational analysis [GOA01a]
Cellular Component: GO:0030288 - outer membrane-bounded periplasmic space
GO:0042597 - periplasmic space Inferred by computational analysis [UniProtGOA11a, UniProtGOA11]
GO:0043190 - ATP-binding cassette (ABC) transporter complex Inferred by computational analysis [GOA01a]

MultiFun Terms: metabolism metabolism of other compounds phosphorous metabolism
transport Channel-type Transporters Pyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active Transporters The ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake Permeases ABC superfamily, periplasmic binding component

Essentiality data for pstS knockouts: ?

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

Credits:
Last-Curated ? 28-Jan-2013 by Mackie A , Macquarie University


Subunit of: 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 [Cox81a]. 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].


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

EC Number: 3.6.3.27

Inhibitors (Competitive): arsenate (Kic = 23µM) [Willsky80]

Kinetic Parameters:

Substrate
Km (μM)
Citations
ATP
23.9
[Sarin01, BRENDA14]


Sequence Features

Feature Class Location Citations Comment
Signal-Sequence 1 -> 25
[Link97, UniProt11]
.
Chain 26 -> 346
[UniProt09]
UniProt: Phosphate-binding protein pstS;
Amino-Acid-Sites-That-Bind 35
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 36
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 63
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 81
[Wang97g, Luecke90]
Forms a very short hydrogen bond with phosphate [Luecke90, Wang97g]. Involved in the discrimination of phosphate from arsenate [Elias12]
Amino-Acid-Sites-That-Bind 160
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 164
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 165
[Luecke90]
Forms hydrogen bond with phosphate
Amino-Acid-Sites-That-Bind 166
[Luecke90]
Forms hydrogen bond with phosphate


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b3728 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10734; confirmed by SwissProt match.


References

Aguena02: Aguena M, Yagil E, Spira B (2002). "Transcriptional analysis of the pst operon of Escherichia coli." Mol Genet Genomics 268(4);518-24. PMID: 12471449

Amemura85: Amemura M, Makino K, Shinagawa H, Kobayashi A, Nakata A (1985). "Nucleotide sequence of the genes involved in phosphate transport and regulation of the phosphate regulon in Escherichia coli." J Mol Biol 184(2);241-50. PMID: 2993631

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." http://www.brenda-enzymes.org.

Byrne14: Byrne RT, Chen SH, Wood EA, Cabot EL, Cox MM (2014). "Surviving extreme exposure to ionizing radiation: Escherichia coli genes and pathways." J Bacteriol. PMID: 25049088

Cox81a: 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

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

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

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

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

Link97: Link AJ, Robison K, Church GM (1997). "Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Electrophoresis 18(8);1259-313. PMID: 9298646

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

Magota84: Magota K, Otsuji N, Miki T, Horiuchi T, Tsunasawa S, Kondo J, Sakiyama F, Amemura M, Morita T, Shinagawa H (1984). "Nucleotide sequence of the phoS gene, the structural gene for the phosphate-binding protein of Escherichia coli." J Bacteriol 157(3);909-17. PMID: 6365894

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

Steed93: Steed PM, Wanner BL (1993). "Use of the rep technique for allele replacement to construct mutants with deletions of the pstSCAB-phoU operon: evidence of a new role for the PhoU protein in the phosphate regulon." J Bacteriol 1993;175(21);6797-809. PMID: 8226621

Surin85: Surin BP, Rosenberg H, Cox GB (1985). "Phosphate-specific transport system of Escherichia coli: nucleotide sequence and gene-polypeptide relationships." J Bacteriol 161(1);189-98. PMID: 3881386

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

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

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

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

Wang94a: 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

Wang97g: 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

Other References Related to Gene Regulation

Aguena08: Aguena M, Ferreira GM, Spira B (2008). "Stability of the pstS transcript of Escherichia coli." Arch Microbiol. PMID: 18820899

Aguena09: Aguena M, Spira B (2009). "Transcriptional processing of the pst operon of Escherichia coli." Curr Microbiol 58(3);264-7. PMID: 19018589

Kimura89: Kimura S, Makino K, Shinagawa H, Amemura M, Nakata A (1989). "Regulation of the phosphate regulon of Escherichia coli: characterization of the promoter of the pstS gene." Mol Gen Genet 1989;215(3);374-80. PMID: 2651888

Makino88: Makino K, Shinagawa H, Amemura M, Kimura S, Nakata A, Ishihama A (1988). "Regulation of the phosphate regulon of Escherichia coli. Activation of pstS transcription by PhoB protein in vitro." J Mol Biol 1988;203(1);85-95. PMID: 3054125

Marzan13: Marzan LW, Hasan CM, Shimizu K (2013). "Effect of acidic condition on the metabolic regulation of Escherichia coli and its phoB mutant." Arch Microbiol 195(3);161-71. PMID: 23274360

Otsuka96: Otsuka J, Watanabe H, Mori KT (1996). "Evolution of transcriptional regulation system through promiscuous coupling of regulatory proteins with operons; suggestion from protein sequence similarities in Escherichia coli." J Theor Biol 1996;178(2);183-204. PMID: 8729576

Salmon03: Salmon K, Hung SP, Mekjian K, Baldi P, Hatfield GW, Gunsalus RP (2003). "Global gene expression profiling in Escherichia coli K12. The effects of oxygen availability and FNR." J Biol Chem 278(32);29837-55. PMID: 12754220

Spira10: Spira B, Aguena M, de Castro Oliveira JV, Yagil E (2010). "Alternative promoters in the pst operon of Escherichia coli." Mol Genet Genomics 284(6);489-98. PMID: 20963440

Spira99: Spira B, Yagil E (1999). "The integration host factor (IHF) affects the expression of the phosphate-binding protein and of alkaline phosphatase in Escherichia coli." Curr Microbiol 38(2);80-5. PMID: 9871104

Surin84: Surin BP, Jans DA, Fimmel AL, Shaw DC, Cox GB, Rosenberg H (1984). "Structural gene for the phosphate-repressible phosphate-binding protein of Escherichia coli has its own promoter: complete nucleotide sequence of the phoS gene." J Bacteriol 1984;157(3);772-8. PMID: 6321434

Taschner04: Taschner NP, Yagil E, Spira B (2004). "A differential effect of sigmaS on the expression of the PHO regulon genes of Escherichia coli." Microbiology 150(Pt 9);2985-92. PMID: 15347756

Taschner06: Taschner NP, Yagil E, Spira B (2006). "The effect of IHF on sigmaS selectivity of the phoA and pst promoters of Escherichia coli." Arch Microbiol 185(3);234-7. PMID: 16404567


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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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