pycA

pyruvate carboxylase

Locus: BSU_14860

Isoelectric point: 5.41

Molecular weight: 127.72 kDa

Protein length: 1148 aa

Gene length: 3447 bp

Function: replenishment of the oxaloacetate pool

Product: pyruvate carboxylase

Essential: no

Synonyms: ylaP

Genomic Context

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Categories containing this gene/protein

• Metabolism → Carbon metabolism → Carbon core metabolism → TCA cycle
• Groups of genes → Membrane proteins

Gene

Coordinates: 1,554,185 1,557,631

The protein

Catalyzed reaction/ biological activity

• ATP + hydrogencarbonate + pyruvate --> ADP + H⁺ + oxaloacetate + phosphate (according to UniProt)

Domains

• ATP-grasp doamin (aa 125-321) (according to UniProt)
• Biotin carboxylation domain (aa 1-457) (according to UniProt)
• Pyruvate carboxyltransferase domain (aa 534-802) (according to UniProt)
• Biotinyl-binding domain (aa 1071-1146) (according to UniProt)

Cofactors

• biotin PubMed

Structure

• 3BG5 (S. aureus) PubMed

Localization

• membrane associated PubMed
• cytoplasm (homogeneously distributed throughout the cell) PubMed

Additional information

• PycA binds to StrepTactin, and may be co-purified when purifying Strep-tagged proteins by SPINE.
• PycA is subject to Clp-dependent proteolysis upon glucose starvation PubMed

Expression and Regulation

Operons

• Genes: ftsW-pycA
  Description: PubMed

  Sigma factors

  • SigA: sigma factor, PubMed, in SigA regulon

  Regulatory mechanism

  • YofA: activation, PubMed, in YofA regulon
  • stringent response: positive regulation, due to presence of adenines at +1 and +2 positions of the transcript PubMed, in stringent response
  • TnrA: repression, in TnrA regulon

  Regulation

  • constitutively expressed PubMed

• Genes: pycA
  Description: PubMed

  Regulation

  • subject to positive stringent control upon lysine starvation PubMed

Biological materials

Mutant

• MGNA-A544 (ylaP::erm), available at the NBRP B. subtilis, Japan
• GP793 (cat), available in Jörg Stülke's lab
• BKE14860 (pycA::erm trpC2) available at BGSC, PubMed, upstream reverse: _UP1_CAAACTTTTCTCCCCCTTAC, downstream forward: _UP4_TAAAAAACAAAGAGTGTATA
• BKK14860 (pycA::kan trpC2) available at BGSC, PubMed, upstream reverse: _UP1_CAAACTTTTCTCCCCCTTAC, downstream forward: _UP4_TAAAAAACAAAGAGTGTATA

Expression vectors

• pGP1289 (N-terminal Strep-tag, purification from B. subtilis, for SPINE, in pGP380), available in Jörg Stülke's lab

References

Reviews

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Original publications

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