Eno

• Description: enolase, glycolytic/ gluconeogenic enzyme, universally conserved protein
  
  

Gene name	eno
Synonyms
Essential	no
Product	enolase
Function	enzyme in glycolysis/ gluconeogenesis
Metabolic function and regulation of this protein in SubtiPathways: Central C-metabolism
MW, pI	46,4 kDa, 4.49
Gene length, protein length	1290 bp, 430 amino acids
Immediate neighbours	yvbK, pgm
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: BSU33900

Phenotypes of a mutant

• no growth on LB, requires glucose and malate
• essential according to Kobayashi et al. on LB PubMed

Database entries

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity: 2-phospho-D-glycerate = phosphoenolpyruvate + H₂O (according to Swiss-Prot) 2-phospho-D-glycerate = phosphoenolpyruvate + H(2)O

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

• Paralogous protein(s):

Extended information on the protein

• Kinetic information: reversible Michaelis-Menten PubMed

• Domains:
  • substrate binding domain (366–369)

• Modification: phosphorylation on Thr-141 AND Ser-259 AND Tyr-281 AND Ser-325 PubMed, PubMed, PubMed

• Cofactor(s): Mg2+

• Effectors of protein activity:
  • Inhibited by EDTA PubMed

• Interactions: Eno-PfkA, Eno-Rny PubMed

• Localization: cytoplasm (according to Swiss-Prot), cytoplasm PubMed and membrane associated PubMed

Database entries

• Structure: 3ES8 (from Oceanobacillus iheyensis, complex with Mg(2+) and malate)

• UniProt: P37869

• KEGG entry: [3]

• E.C. number: 4.2.1.11

Additional information

• Enolase is a moonlighting protein. PubMed
• There are indications that this enzyme is an octamer PubMed
• universally conserved protein

Expression and regulation

• Operon:
  • cggR-gapA-pgk-tpiA-pgm-eno PubMed
  • pgk-tpiA-pgm-eno PubMed

• Sigma factor: SigA PubMed

• Regulation:
  • expression activated by glucose (3.3 fold) PubMed
  • cggR: induced by glycolytic substrates CggR PubMed
  • pgk: constitutive PubMed

• Regulatory mechanism: transcription repression by CggR PubMed

• Additional information:

Biological materials

• Mutant:
• GP594 (eno::cat), available in Stülke lab
• GP599 (eno::erm), available in Stülke lab
• GP698 (eno-pgm::cat), available in Stülke lab

• Expression vector:
  • pGP1426 (expression of eno in B. subtilis, in pBQ200), available in Stülke lab
  • pGP399 (expression of eno from E. coli in B. subtilis, in pBQ200), available in Stülke lab
  • pGP563 (N-terminal His-tag, in pWH844), available in Stülke lab
  • pGP93 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), available in Stülke lab
  • pGP1500 (expression of pgm and eno in B. subtilis, in pBQ200), available in Stülke lab

• lacZ fusion:
  • see pgk

• GFP fusion: pHT315-yfp-eno, available in Mijakovic lab

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

• Antibody: available in Stülke lab

Labs working on this gene/protein

Jörg Stülke, University of Göttingen, Germany Homepage