Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
twitter

Escherichia coli K-12 substr. MG1655 Transporter: putrescine:H+ symporter / putrescine:ornithine antiporter PotE



Gene: potE Accession Numbers: EG10753 (EcoCyc), b0692, ECK0680

Regulation Summary Diagram: ?

Summary:
PotE is a putrescine transporter that is a member of the Amino Acid-Polyamine-Organocation (APC) Superfamily of transporters [Jack00]. In mutational experiments, where all other spermidine and putrescine uptake systems (PotABCD and PotFGHI) were lacking, PotE was found to catalyze uptake of putrescine via a proton symport mechanism with a Km value of 1.8 μM [Kashiwagi97]. In experiments examining PotE function and putrescine uptake in inside-out membrane vesicles, PotE was also found to catalyze putrescine efflux by a putrescine/ornithine antiport system in a 1:1 ratio. The Km for the antiporter activities of putrescine and ornithine were 73 μM and 108 μM, respectively [Kashiwagi97]. Uptake was not disturbed by KCN or CCCP, indicating that the process is not driven by ATP hydrolysis of the proton motive force [Kashiwagi92]. There is also evidence of putrescine/putrescine and ornithine/ornithine antiport. This is supported by experiments where CCCP is used to disable putrescine uptake and the addition of putrescine or ornithine results in rapid efflux of putrescine from the cell [Kashiwagi92]. Hydropathy analysis, PhoA fusions, and B-gal fusions suggest a 12 transmembrane segment topology [Kashiwagi97]. Site directed mutagenesis has implicated the residue E7, 207 or 433 as important for the uptake of putrescine [Kashiwagi97].

Gene Citations: [Kashiwagi91]

Locations: inner membrane

Map Position: [716,169 <- 717,488] (15.44 centisomes)
Length: 1320 bp / 439 aa

Molecular Weight of Polypeptide: 46.495 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0002360 , CGSC:31552 , EchoBASE:EB0746 , EcoGene:EG10753 , EcoliWiki:b0692 , OU-Microarray:b0692 , PortEco:potE , PR:PRO_000023571 , Protein Model Portal:P0AAF1 , RefSeq:NP_415219 , RegulonDB:EG10753 , SMR:P0AAF1 , String:511145.b0692 , UniProt:P0AAF1

Relationship Links: InterPro:IN-FAMILY:IPR002293 , InterPro:IN-FAMILY:IPR004754 , InterPro:IN-FAMILY:IPR027566 , Panther:IN-FAMILY:PTHR11785 , Pfam:IN-FAMILY:PF00324 , Pfam:IN-FAMILY:PF13520

In Paralogous Gene Group: 180 (2 members)

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0015847 - putrescine transport Inferred from experiment Inferred by computational analysis [GOA01, Kashiwagi97]
GO:1903352 - L-ornithine transmembrane transport Inferred by computational analysis Inferred from experiment [Kashiwagi92, GOA01]
GO:0003333 - amino acid transmembrane transport Inferred by computational analysis [GOA01]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11]
GO:0006865 - amino acid transport Inferred by computational analysis [UniProtGOA11, GOA01]
Molecular Function: GO:0015293 - symporter activity Inferred from experiment [Kashiwagi97]
GO:0015496 - putrescine:ornithine antiporter activity Inferred from experiment Inferred by computational analysis [GOA01, Kashiwagi92]
GO:0015171 - amino acid transmembrane transporter activity Inferred by computational analysis [GOA01]
GO:0015297 - antiporter activity Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005886 - plasma membrane Inferred from experiment Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, DiazMejia09, Daley05]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0016021 - integral component of membrane Inferred by computational analysis [UniProtGOA11, GOA01]

MultiFun Terms: cell structure membrane
metabolism carbon utilization amines
metabolism central intermediary metabolism polyamine biosynthesis
transport Electrochemical potential driven transporters Porters (Uni-, Sym- and Antiporters)

Essentiality data for potE 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]

Enzymatic reaction of: putrescine:L-ornithine antiporter (putrescine:H+ symporter / putrescine:ornithine antiporter PotE)

Synonyms: L-ornithine:putrescine antiport

Kinetic Parameters:

Substrate
Km (μM)
Citations
L-ornithine
108.0
[Kashiwagi97]
putrescine
73.0
[Kashiwagi97]


Enzymatic reaction of: putrescine:H+ symporter

Synonyms: Transport of putrescine

Kinetic Parameters:

Substrate
Km (μM)
Citations
putrescine
1.8
[Kashiwagi97]


Sequence Features

Feature Class Location Citations Comment
Transmembrane-Region 11 -> 31
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 38 -> 58
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 90 -> 110
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 117 -> 137
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 152 -> 172
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 186 -> 206
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 224 -> 244
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 277 -> 297
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 325 -> 345
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 352 -> 372
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 387 -> 407
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 410 -> 430
[UniProt10]
UniProt: Helical;; Non-Experimental Qualifier: potential;


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b0692 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10753; confirmed by SwissProt match.


References

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

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

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

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

Jack00: Jack DL, Paulsen IT, Saier MH (2000). "The amino acid/polyamine/organocation (APC) superfamily of transporters specific for amino acids, polyamines and organocations." Microbiology 2000;146 ( Pt 8);1797-814. PMID: 10931886

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

Kashiwagi91: Kashiwagi K, Suzuki T, Suzuki F, Furuchi T, Kobayashi H, Igarashi K (1991). "Coexistence of the genes for putrescine transport protein and ornithine decarboxylase at 16 min on Escherichia coli chromosome." J Biol Chem 1991;266(31);20922-7. PMID: 1939141

Kashiwagi92: Kashiwagi K, Miyamoto S, Suzuki F, Kobayashi H, Igarashi K (1992). "Excretion of putrescine by the putrescine-ornithine antiporter encoded by the potE gene of Escherichia coli." Proc Natl Acad Sci U S A 1992;89(10);4529-33. PMID: 1584788

Kashiwagi97: Kashiwagi K, Shibuya S, Tomitori H, Kuraishi A, Igarashi K (1997). "Excretion and uptake of putrescine by the PotE protein in Escherichia coli." J Biol Chem 1997;272(10);6318-23. PMID: 9045651

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 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."


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Thu Dec 18, 2014, BIOCYC13B.