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

Categories containing this gene/protein

carbon core metabolism

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: part of the RNA degradosome
  • PfkA-Pgm PubMed, PfkA-Eno PubMed
  • PfkA-Rny PubMed, PfkA-PnpA, PfkA-RnjA 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
• extensive information on the structure and enzymatic properties of PfkA can be found at Proteopedia

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 B. subtilis with C-terminal Strep-tag, for SPINE, in pGP382: pGP1266, 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