Eno

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

Gene name	eno
Synonyms
Essential	Yes (PubMed)
Product	enolase
Function	enzyme in glycolysis/ gluconeogenesis
Gene expression levels in SubtiExpress: eno
Interactions involving this protein in SubtInteract: Eno
Metabolic function and regulation of this protein in SubtiPathways: Eno
MW, pI	46,4 kDa, 4.49
Gene length, protein length	1290 bp, 430 amino acids
Immediate neighbours	yvbK, pgm
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, essential genes, membrane proteins, phosphoproteins, universally conserved proteins, most abundant 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

• BsubCyc: BSU33900

• 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

• Cofactors: 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

• BsubCyc: BSU33900

• 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:
  • belongs to the 100 most abundant proteins PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium): 10563 PubMed
  • number of protein molecules per cell (complex medium with amino acids, without glucose): 9957 PubMed

Biological materials

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

• Expression vector:
  • pGP1426 (expression of eno in B. subtilis, in pBQ200), available in Jörg Stülke's lab
  • pGP1500 (expression of pgm and eno in B. subtilis, in pBQ200), available in Jörg Stülke's lab
  • pGP563 (N-terminal His-tag, in pWH844), available in Jörg Stülke's lab
  • pGP1276 (N-terminal Strep-tag, purification from E. coli, in pGP172), available in Jörg Stülke's lab
  • pGP93 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), available in Jörg Stülke's lab
  • GP1215 (chromosomal eno-Strep fusion, spc), purification from B. subtilis, for SPINE, available in Jörg Stülke's 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 Jörg Stülke's lab

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

• Antibody: available in Jörg Stülke's 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)