PckA

• Description: phosphoenolpyruvate carboxykinase
  
  

Gene name	pckA
Synonyms	ppc
Essential	no
Product	phosphoenolpyruvate carboxykinase
Function	synthesis of phosphoenolpyruvate
Metabolic function and regulation of this protein in SubtiPathways: Central C-metabolism
MW, pI	58,1 kDa, 5.12
Gene length, protein length	1581 bp, 527 amino acids
Immediate neighbours	metK, ytmB
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: BSU30560

Database entries

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity: ATP + oxaloacetate = ADP + phosphoenolpyruvate + CO₂ (according to Swiss-Prot) ATP + oxaloacetate = ADP + phosphoenolpyruvate + CO(2)

• Protein family: phosphoenolpyruvate carboxykinase [ATP] family (according to Swiss-Prot) phosphoenolpyruvate carboxykinase [ATP] family

• Paralogous protein(s):

Extended information on the protein

• Kinetic information:

• Domains:
  • Nucleotide binding Domain (233–240)

• Modification:

• Cofactor(s):

• Effectors of protein activity:

• Interactions:
  • Mdh-PckA (this interaction occurs under gluconeogenic, but not under glycolytic conditions) PubMed

• Localization: cytoplasm (according to Swiss-Prot), cytoplasm

Database entries

• Structure: 2PXZ (E.coli)

• UniProt: P54418

• KEGG entry: [3]

• E.C. number: 4.1.1.49

Expression and regulation

• Operon: pckA PubMed

• Sigma factor: SigA PubMed

• Regulation:
  • repressed by glucose (35-fold) CcpN PubMed

• Regulatory mechanism:
  • CcpN: transcription repression PubMed

• Additional information:

Biological materials

• Mutant:

• Expression vector:
  • pGP1753 (N-terminal Strep-tag, for SPINE, purification from B. subtilis, in pGP380) (available in Stülke lab)
  • pGP1762 (for expression, purification in E. coli with N-terminal His-tag, in pWH844, available in Stülke lab)
  • pGP1763 (for expression, purification in E. coli with N-terminal Strep-tag in pGP172, available in Stülke lab)

• lacZ fusion:

• GFP fusion:

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

• Antibody:

• FLAG-tag construct: GP1102 (spc, based on pGP1331), available in the Stülke lab

Labs working on this gene/protein

Stephane Aymerich, Microbiology and Molecular Genetics, INRA Paris-Grignon, France