PckA

• Description: phosphoenolpyruvate carboxykinase
  
  

Gene name	pckA
Synonyms	ppc
Essential	no
Product	phosphoenolpyruvate carboxykinase
Function	synthesis of phosphoenolpyruvate
Gene expression levels in SubtiExpress: pckA
Interactions involving this protein in SubtInteract: PckA
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
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

This gene is a member of the following regulons

CcpN regulon

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

• Expression browser: 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:
  • 1A1005 ( pckA::spec), PubMed, available at BGSC
  • 1A996 ( pckA::spec), PubMed, available at BGSC
  • GP1147 (pckA::neo), available in Jörg Stülke's lab

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

• lacZ fusion:

• GFP fusion: GP1430 (spc, based on pGP1870), available in the Stülke lab

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

• Antibody:

• FLAG-tag construct:
  • GP1129 (spc, based on pGP1331), available in Jörg Stülke's lab

Labs working on this gene/protein

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