pckA

168

phosphoenolpyruvate carboxykinase

locus: BSU_30560

Molecular weight: 58.13 kDa

pI: 5.12

Protein length: 527 aa

Gene length: 1584 bp

function: synthesis of phosphoenolpyruvate

product: phosphoenolpyruvate carboxykinase

essential: no

ec: 4.1.1.49

synonyms: pckA

List of homologs in different organisms, belongs to COG1866 (Galperin et al., 2021)

Gene

Coordinates: 3,129,530  3,131,113

The protein

Catalyzed reaction/ biological activity

• [metabolite|ATP] + [metabolite|oxaloacetate] --> [metabolite|ADP] + CO₂ + [metabolite|phosphoenolpyruvate] (according to UniProt)

Protein family

• phosphoenolpyruvate carboxykinase (ATP) family (single member, according to UniProt)

[wiki|Domains]

• Nucleotide binding Domain (233240)

Structure

• [PDB|1J3B] (from ''Thermus thermophilus'', 55.1% identity) [pubmed|16239727]
• [AF|P54418]

[wiki|Localization]

• cytoplasm (according to Swiss-Prot),  cytoplasm

Additional information

• [protein|74D4616D1A550D87182FAAC2A4AF6E04F81E1572|pckA] is degraded in the forespore using [protein|7CFDFB42AF816FF5766086E537472008E6CB7E85|mdfA] as the adaptor protein [pubmed|39651166]

Expression and Regulation

Operons

• genes: [gene|74D4616D1A550D87182FAAC2A4AF6E04F81E1572|pckA]

  description: [Pubmed|9387221]

  regulation

  • repressed by glucose (35-fold) [protein|search|CcpN] [Pubmed|15720552,12850135]

  regulatory mechanism

  • [protein|2234D81F8F9B92EAF1CF6F73BEC92D4DE54E2636|ccpN]: repression, [Pubmed|15720552], in [regulon|protein:2234D81F8F9B92EAF1CF6F73BEC92D4DE54E2636|ccpN regulon]

  sigma factors

  • [protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA]: sigma factor, [Pubmed|15720552], in [regulon|protein:360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA regulon]

Biological materials

Mutant

• 1A1005 ( ''pckA''::''spec''), [Pubmed| ], available at [http://pasture.asc.ohio-state.edu/BGSC/getdetail.cfm?bgscid=1A1005&Search=1A1005 BGSC]
• 1A996 ( ''pckA''::''spec''), [Pubmed| ], available at [http://pasture.asc.ohio-state.edu/BGSC/getdetail.cfm?bgscid=1A996&Search=1A996 BGSC]
• GP1147 (''pckA''::''neo''), available in [wiki|Jörg Stülke]'s lab
• BKE30560 ([gene|74D4616D1A550D87182FAAC2A4AF6E04F81E1572|pckA]::erm  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKE30560 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CATATGAAACCTTCCTTTAT,  downstream forward: _UP4_TAAAAAACAAAAGCCAAGAG
• BKK30560 ([gene|74D4616D1A550D87182FAAC2A4AF6E04F81E1572|pckA]::kan  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKK30560 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CATATGAAACCTTCCTTTAT,  downstream forward: _UP4_TAAAAAACAAAAGCCAAGAG

Expression vectors

• pGP1753 (N-terminal Strep-tag, for [wiki|SPINE], purification from ''B. subtilis'', in [wiki|pGP380]) (available in [wiki|Jörg Stülke]'s lab) [pubmed|20933603]
• pGP1762 (for expression, purification in ''E. coli'' with N-terminal His-tag, in [wiki|pWH844], available in [wiki|Jörg Stülke]'s lab)
• pGP1763 (for expression, purification in ''E. coli'' with N-terminal Strep-tag, in [wiki|pGP172], available in [wiki|Jörg Stülke]'s lab)

two-hybrid system

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

FLAG-tag construct

• GP1129 (spc, based on [wiki|pGP1331]), available in [wiki|Jörg Stülke]'s lab [pubmed|20933603]

lacZ fusion

• pGP3312 (cat, based on [wiki|pAC5]), available in [wiki|Jörg Stülke]'s lab

GFP fusion

• GP1430 (spc, based on [wiki|pGP1870]), available in [wiki|Jörg Stülke]'s lab

labs

• [wiki|Stephane Aymerich], Microbiology and Molecular Genetics, INRA Paris-Grignon, France

References

• 36815589,12850135,15720552,18586936,9387221,20933603,22190493,23136871,39651166,16239727