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Escherichia coli K-12 substr. MG1655 Polypeptide: anaerobic glycerol-3-phosphate dehydrogenase subunit A



Gene: glpA Accession Numbers: EG10391 (EcoCyc), b2241, ECK2233

Regulation Summary Diagram: ?

Component of: glycerol-3-phosphate dehydrogenase, anaerobic (extended summary available)

Summary:
GlpA is the large subunit of a three subunit glycerol-3-phosphate dehydrogenase complex. In anaerobic conditions this respiratory enzyme converts glycerol-3-phosphate to dihydroxyacetone phosphate (DHAP) using fumarate as a terminal electron acceptor. The GlpA subunit contains noncovalently bound FAD. [Cole88, Schryvers82, Schryvers81, Kistler71].

Citations: [Iuchi90, Ehrmann87, Miki79a, Freedberg73]

Gene Citations: [Larson92, Ehrmann87]

Locations: cytosol, inner membrane

Map Position: [2,350,669 -> 2,352,297] (50.66 centisomes)
Length: 1629 bp / 542 aa

Molecular Weight of Polypeptide: 58.958 kD (from nucleotide sequence), 62.0 kD (experimental) [Schryvers81 ]

Unification Links: ASAP:ABE-0007422 , CGSC:694 , EchoBASE:EB0386 , EcoGene:EG10391 , EcoliWiki:b2241 , ModBase:P0A9C0 , OU-Microarray:b2241 , PortEco:glpA , PR:PRO_000022792 , Pride:P0A9C0 , Protein Model Portal:P0A9C0 , RefSeq:NP_416744 , RegulonDB:EG10391 , SMR:P0A9C0 , String:511145.b2241 , UniProt:P0A9C0

Relationship Links: InterPro:IN-FAMILY:IPR000447 , InterPro:IN-FAMILY:IPR006076 , InterPro:IN-FAMILY:IPR007419 , InterPro:IN-FAMILY:IPR017752 , Pfam:IN-FAMILY:PF01266 , Pfam:IN-FAMILY:PF04324 , Prints:IN-FAMILY:PR01001 , Prosite:IN-FAMILY:PS00977 , Prosite:IN-FAMILY:PS00978

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0009061 - anaerobic respiration Inferred from experiment [Kuritzkes84]
GO:0046168 - glycerol-3-phosphate catabolic process Inferred from experiment [Varga95, Schryvers81]
GO:0006072 - glycerol-3-phosphate metabolic process Inferred by computational analysis [GOA01a]
GO:0019563 - glycerol catabolic process Inferred by computational analysis [UniProtGOA12]
GO:0046174 - polyol catabolic process Inferred by computational analysis [GOA01a]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11a, GOA01a]
Molecular Function: GO:0004368 - glycerol-3-phosphate dehydrogenase activity Inferred from experiment Inferred by computational analysis [GOA01a, Varga95, Kistler71, Schryvers81]
GO:0005515 - protein binding Inferred from experiment [Schryvers81]
GO:0050660 - flavin adenine dinucleotide binding Inferred from experiment Inferred by computational analysis [GOA01a, Cole88, Schryvers81]
GO:0010181 - FMN binding Inferred by computational analysis [GOA01a]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11a, GOA01a]
GO:0052591 - sn-glycerol-3-phosphate:ubiquinone-8 oxidoreductase activity Inferred by computational analysis [GOA01]
Cellular Component: GO:0005829 - cytosol Inferred from experiment [Schryvers82]
GO:0009331 - glycerol-3-phosphate dehydrogenase complex Inferred from experiment Inferred by computational analysis [GOA01a, Varga95]
GO:0005886 - plasma membrane Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA01a]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a]

MultiFun Terms: cell structure membrane
metabolism central intermediary metabolism misc. glycerol metabolism
metabolism energy metabolism, carbon anaerobic respiration
metabolism energy production/transport electron donors

Essentiality data for glpA 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-Oct-2013 by Kubo A , SRI International


Subunit of: glycerol-3-phosphate dehydrogenase, anaerobic

Synonyms: G3P dehydrogenase, sn-glycerol-3-phosphate:(acceptor) 2-oxidoreductase

Subunit composition of glycerol-3-phosphate dehydrogenase, anaerobic = [GlpA][GlpB][GlpC]
         anaerobic glycerol-3-phosphate dehydrogenase subunit A = GlpA (summary available)
         anaerobic glycerol-3-phosphate dehydrogenase subunit B = GlpB (summary available)
         anaerobic glycerol-3-phosphate dehydrogenase subunit C = GlpC (summary available)

Summary:
glpABC encodes anaerobic glycerol-3-phosphate dehydrogenase which catalyses the oxidation of glycerol-3-phosphate to dihyroxyacetone phosphate. GlpABC is a respiratory enzyme; anaerobic growth of E. coli with glycerol and fumarate induces expression of an anaerobic glycerol-3-phosphate dehydrogenase and fumarate reductase and is associated with proton translocation and the generation of a proton motive force [Miki78, Miki73, Kistler71].

The GlpABC enzyme is loosely associated with the cell membrane. A functional two subunit form, GlpAB, has been isolated and characterised; it is assumed that the third subunit (GlpC) is responsible for membrane anchoring. The GlpAB complex contains contains 1 molecule of FAD and 2 non-haem irons per dimer; the GlpB subunit is thought to bind flavin mononucleotide [Schryvers81, Cole88]. The GlpC subunit contains two iron-sulfur binding sites; it does not contain any transmembrane helices, so the mechanism by which it acts as the membrane anchor for the complex is not clear [Cole88, Varga95]. Overexpressed GlpC associates with the inner membrane. GlpC is a participant in the electron transport path ([Varga95] - note that this paper uses the name GlpB for the protein encoded by the third gene in the glp operon).

glpABC is subject to complex transcriptional regulation. Expression of glpABC (along with other members of the glp regulon) is repressed by GlpR and induced by glycerol-3-phosphate; the glp regulon is subject to catabolite repression during growth in the presence of glucose (reviewed in [Lin76], [Larson92] ). Expression of glpABC is positively regulated by the the Fnr transcription regulator [Kuritzkes84, Iuchi90].

E. coli K-12 contains two glycerol-3-phosphate dehydrogenases encoded by the glpABC and glpD genes. GlpABC is required for anaerobic growth with glycerol or glycerol-3-phosphate and fumarate as the terminal electron acceptor while GlpD is required for aerobic growth with glycerol (or glycerol-3-phosphate). Either enzyme suffices for anaerobic growth on glycerol and nitrate ([Kistler71, Miki75] and review by [Stewart88]. The ratio of GlpD to GlpABC is high when oxygen or nitrate serves as a terminal electron acceptor and low when fumarate serves this role [Freedberg73].

Citations: [Yang97]

Locations [Comment 5]: cytosol

GO Terms:

Biological Process: GO:0009061 - anaerobic respiration Inferred from experiment [Kistler71, Miki78]
GO:0046168 - glycerol-3-phosphate catabolic process Inferred from experiment [Schryvers81]
Molecular Function: GO:0004368 - glycerol-3-phosphate dehydrogenase activity Inferred from experiment [Schryvers81]
GO:0050660 - flavin adenine dinucleotide binding Inferred from experiment [Schryvers81]
Cellular Component: GO:0009331 - glycerol-3-phosphate dehydrogenase complex Inferred from experiment [Varga95]

Credits:
Last-Curated ? 06-Aug-2014 by Mackie A , Macquarie University


Enzymatic reaction of: glycerol-3-phosphate dehydrogenase, anaerobic

Synonyms: G3P dehydrogenase, sn-glycerol-3-phosphate:(acceptor) 2-oxidoreductase

EC Number: 1.1.5.3

In Pathways: glycerol and glycerophosphodiester degradation , glycerol degradation I , glycerophosphodiester degradation , glycerol-3-phosphate to fumarate electron transfer

Cofactors or Prosthetic Groups: FAD [Schryvers81], FMN [Schryvers81]

Inhibitors (Unknown Mechanism): p-chloromercuriphenylsulfonate [Schryvers81] , 5,5'-dithio-bis-2-nitrobenzoate [Schryvers81]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
sn-glycerol 3-phosphate
100.0, 339.0
33.3
[Schryvers81, BRENDA14]

T(opt): 44 °C [BRENDA14, Schryvers78]

pH(opt): 7 [BRENDA14, Schryvers78]


Sequence Features

Feature Class Location Citations Comment
Nucleotide-Phosphate-Binding-Region 10 -> 38
[UniProt10]
UniProt: FAD; Non-Experimental Qualifier: potential;
Sequence-Conflict 329
[Cole88, UniProt10a]
Alternate sequence: V → L; UniProt: (in Ref. 1; AAA83864);


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b2241 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10391; 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

Bairoch93: Bairoch A, Boeckmann B (1993). "The SWISS-PROT protein sequence data bank, recent developments." Nucleic Acids Res. 21:3093-3096. PMID: 8332529

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Cole88: Cole ST, Eiglmeier K, Ahmed S, Honore N, Elmes L, Anderson WF, Weiner JH (1988). "Nucleotide sequence and gene-polypeptide relationships of the glpABC operon encoding the anaerobic sn-glycerol-3-phosphate dehydrogenase of Escherichia coli K-12." J Bacteriol 1988;170(6);2448-56. PMID: 3286606

Ehrmann87: Ehrmann M, Boos W, Ormseth E, Schweizer H, Larson TJ (1987). "Divergent transcription of the sn-glycerol-3-phosphate active transport (glpT) and anaerobic sn-glycerol-3-phosphate dehydrogenase (glpA glpC glpB) genes of Escherichia coli K-12." J Bacteriol 169(2);526-32. PMID: 3027032

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

Freedberg73: Freedberg WB, Lin EC (1973). "Three kinds of controls affecting the expression of the glp regulon in Escherichia coli." J Bacteriol 115(3);816-23. PMID: 4580569

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."

Iuchi90: Iuchi S, Cole ST, Lin EC (1990). "Multiple regulatory elements for the glpA operon encoding anaerobic glycerol-3-phosphate dehydrogenase and the glpD operon encoding aerobic glycerol-3-phosphate dehydrogenase in Escherichia coli: further characterization of respiratory control." J Bacteriol 1990;172(1);179-84. PMID: 2403539

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

Kistler71: Kistler WS, Lin EC (1971). "Anaerobic L- -glycerophosphate dehydrogenase of Escherichia coli: its genetic locus and its physiological role." J Bacteriol 108(3);1224-34. PMID: 4945192

Kuritzkes84: Kuritzkes DR, Zhang XY, Lin EC (1984). "Use of phi(glp-lac) in studies of respiratory regulation of the Escherichia coli anaerobic sn-glycerol-3-phosphate dehydrogenase genes (glpAB)." J Bacteriol 157(2);591-8. PMID: 6363389

Larson92: Larson TJ, Cantwell JS, van Loo-Bhattacharya AT (1992). "Interaction at a distance between multiple operators controls the adjacent, divergently transcribed glpTQ-glpACB operons of Escherichia coli K-12." J Biol Chem 1992;267(9);6114-21. PMID: 1556120

Lin76: Lin EC (1976). "Glycerol dissimilation and its regulation in bacteria." Annu Rev Microbiol 1976;30;535-78. PMID: 825019

Miki73: Miki K, Lin EC (1973). "Enzyme complex which couples glycerol-3-phosphate dehydrogenation to fumarate reduction in Escherichia coli." J Bacteriol 114(2);767-71. PMID: 4574699

Miki75: Miki K, Lin EC (1975). "Electron transport chain from glycerol 3-phosphate to nitrate in Escherichia coli." J Bacteriol 124(3);1288-94. PMID: 127786

Miki78: Miki K, Wilson TH (1978). "Proton translocation associated with anaerobic transhydrogenation from glycerol 3-phosphate to fumarate in Escherichia coli." Biochem Biophys Res Commun 83(4);1570-5. PMID: 29636

Miki79a: Miki K, Silhavy TJ, Andrews KJ (1979). "Resolution of glpA and glpT loci into separate operons in Escherichia coli K-12 strains." J Bacteriol 138(1);268-9. PMID: 374371

Schryvers78: Schryvers A, Lohmeier E, Weiner JH (1978). "Chemical and functional properties of the native and reconstituted forms of the membrane-bound, aerobic glycerol-3-phosphate dehydrogenase of Escherichia coli." J Biol Chem 253(3);783-8. PMID: 340460

Schryvers81: Schryvers A, Weiner JH (1981). "The anaerobic sn-glycerol-3-phosphate dehydrogenase of Escherichia coli. Purification and characterization." J Biol Chem 1981;256(19);9959-65. PMID: 6792201

Schryvers82: Schryvers A, Weiner JH (1982). "The anaerobic sn-glycerol-3-phosphate dehydrogenase: cloning and expression of the glpA gene of Escherichia coli and identification of the glpA products." Can J Biochem 60(3);224-31. PMID: 6282417

Stewart88: Stewart V (1988). "Nitrate respiration in relation to facultative metabolism in enterobacteria." Microbiol Rev 52(2);190-232. PMID: 3045516

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

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."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Varga95: Varga ME, Weiner JH (1995). "Physiological role of GlpB of anaerobic glycerol-3-phosphate dehydrogenase of Escherichia coli." Biochem Cell Biol 73(3-4);147-53. PMID: 7576488

Yang97: Yang B, Gerhardt SG, Larson TJ (1997). "Action at a distance for glp repressor control of glpTQ transcription in Escherichia coli K-12." Mol Microbiol 24(3);511-21. PMID: 9179845

Other References Related to Gene Regulation

Bradley07: Bradley MD, Beach MB, de Koning AP, Pratt TS, Osuna R (2007). "Effects of Fis on Escherichia coli gene expression during different growth stages." Microbiology 153(Pt 9);2922-40. PMID: 17768236

Pruss01: Pruss BM, Liu X, Hendrickson W, Matsumura P (2001). "FlhD/FlhC-regulated promoters analyzed by gene array and lacZ gene fusions." FEMS Microbiol Lett 2001;197(1);91-7. PMID: 11287152

Weissenborn92: Weissenborn DL, Wittekindt N, Larson TJ (1992). "Structure and regulation of the glpFK operon encoding glycerol diffusion facilitator and glycerol kinase of Escherichia coli K-12." J Biol Chem 1992;267(9);6122-31. PMID: 1372899

Wong92: Wong KK, Kwan HS (1992). "Transcription of glpT of Escherichia coli K12 is regulated by anaerobiosis and fnr." FEMS Microbiol Lett 1992;73(1-2);15-8. PMID: 1521763

Zhao94: Zhao N, Oh W, Trybul D, Thrasher KS, Kingsbury TJ, Larson TJ (1994). "Characterization of the interaction of the glp repressor of Escherichia coli K-12 with single and tandem glp operator variants." J Bacteriol 176(8);2393-7. PMID: 8157609

Zheng04: Zheng D, Constantinidou C, Hobman JL, Minchin SD (2004). "Identification of the CRP regulon using in vitro and in vivo transcriptional profiling." Nucleic Acids Res 32(19);5874-93. PMID: 15520470


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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 Nov 27, 2014, biocyc12.