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: pfkA
MW, pI	34,1 kDa, 6.14
Gene length, protein length	957 bp, 319 amino acids
Immediate neighbours	pyk, accA
Sequences	Protein DNA DNA_with_flanks
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 according to PubMed, non-essential according to PubMed

Database entries

• BsubCyc: BSU29190

• 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

• BsubCyc: BSU29190

• 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, PubMed
• GP595 (pfkA::erm), available in Jörg Stülke's lab, PubMed

• 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