PfkA

• Description: phosphofructokinase, glycolytic enzyme
  
  

Gene name	pfkA
Synonyms	pfk
Essential	no
Product	6-phosphofructokinase
Function	catabolic enzyme in glycolysis
Gene expression levels in SubtiExpress: pfkA
Interactions involving this protein in SubtInteract: PfkA
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
Expression at a glance   PubMed

Categories containing this gene/protein

carbon core metabolism, phosphoproteins

The gene

Basic information

• Locus tag: BSU29190

Phenotypes of a mutant

• essential PubMed, non-essential according to PubMed

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:
  • phosphorylated on Arg-233 PubMed

• 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:
  • forms homotetramers PubMed
  • part of the RNA degradosome PubMed
  • PfkA-Pgm PubMed
  • PfkA-Eno PubMed, K(D) of the interaction: 40 nM PubMed
  • PfkA-Rny PubMed
  • PfkA-PnpA PubMed
  • PfkA-RnjA PubMed,

• Localization: cytoplasm (Homogeneous) PubMed

Database entries

• Structure:
  • 4A3S PubMed
  • 1MTO (mutant, complex with fructose-6-phosphate, Geobacillus stearothermophilus)
  • 4PFK (Geobacillus stearothermophilus)

• 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

• Expression browser: pfkA PubMed

• Sigma factor:

• Regulation:
  • twofold induced by glucose PubMed

• Regulatory mechanism:

• Additional information:

Biological materials

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

• Expression vector:
  • for expression/ purification from B. subtilis with N-terminal Strep-tag, for SPINE, in pGP380: pGP87, available in Jörg Stülke's lab
  • for expression/ purification from B. subtilis with C-terminal Strep-tag, for SPINE, in pGP382: pGP1266, available in Jörg Stülke's lab
  • for expression/ purification from E. coli with N-terminal His-tag, in pWH844: pGP393, available in Jörg Stülke's lab
  • for expression in B. subtilis, in pBQ200: pGP1422, available in Jörg Stülke's lab

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

• GFP fusion:

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

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

• Antibody:

Labs working on this gene/protein

Jörg Stülke, University of Göttingen, Germany Homepage