Testing

• Description: enolase, glycolytic/ gluconeogenic enzyme
  
  

Gene name	eno
Synonyms
Essential	yes
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	pgm, yvgK
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

• Essential 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)

• Swiss prot entry: P37869

• KEGG entry: [3]

• E.C. number: 4.2.1.11

Additional information

There are indications that this enzyme is an octamer PubMed

Expression and regulation

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

• Sigma factor: SigA

• Regulation: expression activated by glucose (3.3 fold) PubMed
  • cggR: neg. regulated by CggR PubMed, induced by sugar
  • pgk: constitutive PubMed

• Regulatory mechanism: transcription repression by CggR PubMed

• Additional information:

Biological materials

• Mutant:

• Expression vector: pGP563 (N-terminal His-tag, in pWH844), pGP93 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), available in Stülke lab

• lacZ fusion:

• 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