Sandbox

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

Gene name	eno
Synonyms
Essential	no
Product	enolase
Function	enzyme in glycolysis/ gluconeogenesis
Interactions involving this protein in SubtInteract: Eno
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
Expression

Categories containing this gene/protein

carbon core metabolism, membrane proteins, phosphoproteins, universally conserved proteins

This gene is a member of the following regulons

CggR regulon

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:
  • forms octamers PubMed
  • part of the RNA degradosome PubMed
  • Eno-PfkA PubMed, K(D) of the interaction: 40 nM PubMed
  • Eno-Rny PubMed, K(D) of the interaction: 100 nM PubMed
  • Eno-CshA PubMed

• Localization:
  • cytoplasm PubMed
  • membrane associated PubMed
  • exported, this requires a long, unbent α-helix (from A108 to L126) PubMed

Database entries

• Structure:
  • 4A3R PubMed
  • 1W6T (from Streptococcus pneumoniae) PubMed

• 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
• extensive information on the structure and enzymatic properties of Eno can be found at Proteopedia

Expression and regulation

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

• Expression browser: 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
  • pGP1500 (expression of pgm and eno in B. subtilis, in pBQ200), available in Stülke lab
  • pGP563 (N-terminal His-tag, in pWH844), available in Stülke lab
  • pGP1276 (N-terminal Strep-tag, purification from E. coli, in pGP172), available in Stülke lab
  • pGP93 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), available in Stülke lab
  • GP1215 (eno-Strep (spc)), purification from B. subtilis, for SPINE, 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

• FLAG-tag construct: GP1214 (spc, based on pGP1331), available in the Stülke lab

• Antibody: available in Stülke lab

Labs working on this gene/protein

Jörg Stülke, University of Göttingen, Germany Homepage

References

Reviews

G H Reed, R R Poyner, T M Larsen, J E Wedekind, I Rayment
Structural and mechanistic studies of enolase.
Curr Opin Struct Biol: 1996, 6(6);736-43
[PubMed:8994873] [WorldCat.org] [DOI] (P p)

Subcellular localization of enolase

Additional publications: PubMed