PfkA

• Description: phosphofructokinase, glycolytic enzyme
  
  

Gene name	pfkA
Synonyms	pfk
Essential	no
Product	6-phosphofructokinase
Function	catabolic enzyme in glycolysis
Metabolic function and regulation of this protein in SubtiPathways: Central C-metabolism, Sugar catabolism
MW, pI	34,1 kDa, 6.14
Gene length, protein length	957 bp, 319 amino acids
Immediate neighbours	pyk, accA
Get the DNA and protein sequences (Barbe et al., 2009)
Genetic context This image was kindly provided by SubtiList

The gene

Basic information

• Locus tag: BSU29190

Phenotypes of a mutant

• essential according to Kobayashi et al., dispensable according to Munoz-Marquez and Ponce-Rivas

Database entries

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity: ATP + D-fructose 6-phosphate = ADP + D-fructose 1,6-bisphosphate (according to Swiss-Prot)

• Protein family: phosphofructokinase family (according to Swiss-Prot) phosphofructokinase family

• Paralogous protein(s):

Extended information on the protein

• Kinetic information: Allosteric Regulation (Reversible) PubMed

• Domains:
  • 3 x nucleotide binding domain (ATP) (21–25), (154–158), (171–187)

• Modification:

• Cofactor(s): Mg2+

• Effectors of protein activity:
  • Inhibited by citrate, PEP (Hill Coefficient 3) and Ca2+ (competes with Mg2+) in B. licheniformes PubMed.
  • Inhibited by ATP (competitively) and f6p (non-competitively) in G. stearothermophillus PubMed
  • Activated by GDP and ADP in lower concentrations (1mM); above that inhibition, competing with the ATP for the binding site (in G. stearothermophillus) PubMed
  • Activated by NH4+ PubMed

• Interactions: PfkA-Pgm PubMed, PfkA-Eno PubMed, PfkA-Rny PubMed, PfkA-PnpA, PfkA-RnjA PubMed, Eno-CshA PubMed

• Localization: cytoplasm (Homogeneous) PubMed

Database entries

• Structure: 1MTO (mutant, complex with fructose-6-phosphate, Geobacillus stearothermophilus), 4PFK (Geobacillus stearothermophilus), Geobacillus stearothermophilus NCBI, Mutant form, complex with fructose-6-phosphate Geobacillus stearothermophilus NCBI

• UniProt: O34529

• KEGG entry: [3]

• E.C. number: 2.7.1.11

Additional information

PfkA is a moonlighting protein. PubMed

Expression and regulation

• Operon: pfkA-pyk-ytzA PubMed

• Sigma factor:

• Regulation:
  • twofold induced by glucose PubMed

• Regulatory mechanism:

• Additional information:

Biological materials

• Mutant:
• GP590 (pfkA::cat), available in Stülke lab
• GP595 (pfkA::erm), available in Stülke lab

• Expression vector:
  • for expression/ purification from B. subtilis with N-terminal Strep-tag, for SPINE, in pGP380: pGP87, available in Stülke lab
  • for expression/ purification from E. coli with N-terminal His-tag, in pWH844: pGP393, available in Stülke lab
  • for expression in B. subtilis, in pBQ200: pGP1422, available in Stülke lab

• lacZ fusion: pGP511 (in pAC6), available in Stülke lab

• GFP fusion:

• two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Stülke lab

• FLAG-tag construct: GP1019 (spc, based on pGP1331), available in the Stülke lab

• Antibody:

Labs working on this gene/protein

Jörg Stülke, University of Göttingen, Germany Homepage

References

Martin Lehnik-Habrink, Henrike Pförtner, Leonie Rempeters, Nico Pietack, Christina Herzberg, Jörg Stülke
The RNA degradosome in Bacillus subtilis: identification of CshA as the major RNA helicase in the multiprotein complex.
Mol Microbiol: 2010, 77(4);958-71
[PubMed:20572937] [WorldCat.org] [DOI] (I p)

María-Enriqueta Muñoz-Márquez, Elizabeth Ponce-Rivas
Effect of pfkA chromosomal interruption on growth, sporulation, and production of organic acids in Bacillus subtilis.
J Basic Microbiol: 2010, 50(3);232-40
[PubMed:20473954] [WorldCat.org] [DOI] (I p)

Fabian M Commichau, Fabian M Rothe, Christina Herzberg, Eva Wagner, Daniel Hellwig, Martin Lehnik-Habrink, Elke Hammer, Uwe Völker, Jörg Stülke
Novel activities of glycolytic enzymes in Bacillus subtilis: interactions with essential proteins involved in mRNA processing.
Mol Cell Proteomics: 2009, 8(6);1350-60
[PubMed:19193632] [WorldCat.org] [DOI] (I p)

Jean-Christophe Meile, Ling Juan Wu, S Dusko Ehrlich, Jeff Errington, Philippe Noirot
Systematic localisation of proteins fused to the green fluorescent protein in Bacillus subtilis: identification of new proteins at the DNA replication factory.
Proteomics: 2006, 6(7);2135-46
[PubMed:16479537] [WorldCat.org] [DOI] (P p)

H Ludwig, G Homuth, M Schmalisch, F M Dyka, M Hecker, J Stülke
Transcription of glycolytic genes and operons in Bacillus subtilis: evidence for the presence of multiple levels of control of the gapA operon.
Mol Microbiol: 2001, 41(2);409-22
[PubMed:11489127] [WorldCat.org] [DOI] (P p)

X Zhu, M Byrnes, J W Nelson, S H Chang
Role of glycine 212 in the allosteric behavior of phosphofructokinase from Bacillus stearothermophilus.
Biochemistry: 1995, 34(8);2560-5
[PubMed:7873536] [WorldCat.org] [DOI] (P p)

M Byrnes, X Zhu, E S Younathan, S H Chang
Kinetic characteristics of phosphofructokinase from Bacillus stearothermophilus: MgATP nonallosterically inhibits the enzyme.
Biochemistry: 1994, 33(11);3424-31
[PubMed:8136379] [WorldCat.org] [DOI] (P p)

C K Marschke, R W Bernlohr
Purification and characterization of phosphofructokinase of Bacillus licheniformis.
Arch Biochem Biophys: 1973, 156(1);1-16
[PubMed:4269800] [WorldCat.org] [DOI] (P p)